genomic instability in cancer and aging

Genomic instability in cancer and agingJan Vijg, University of Texas Health Science Center, San Antonio, Texas

DNA mutations

Role of genome instability in agingEvolutionary logic of genome instabilityEvidence for genome deterioration in agingAssociation of DNA repair defects with accelerated aging

Effect of deleterious mutations on fitness in E. coliElena & Lenski, 1997, Nature 390: 395

The problem in this theory is that of developing a quantitative measure of mutations in somatic cells

Howard J. Curtis, 1963

LacZ plasmid recovery

Spontaneous mutant frequencies with age in heart and small intestine

Spontaneous mutant frequencies with age in mouse organs

Mutant spectra in vivo and in vitro

Point mutational spectra in organs from Young (3m) and Old (32m) miceMutant frequency (x10-5)PercentageBrainHeartLiverSpleenSmall intestineMock recoveryLymphomaMutant frequency (x10-5)PercentageOY

Mutant frequencies in vivo and in vitro

MEF 6Growth curveMutant frequencyspectra3%20%Total MFPoint MutSize change

Chart2

00

1.32048708091.3204870809

3.62985732253.3407818431

5.84605347784.9668315359

7.99769585245.7037971301

9.57300818317.042934515

11.32003746068.0286077335

12.83941161979.0209082086

14.547230868213.8868270232

17.664925910916.9537772671

20.4212375939

Embryo 6- 3% O2

Embryo 6-20% O2

Days

Population Doublings

Sheet3

Cell typeDateDay# totalPD#total

Embryo 5-3% O223-May-9800

Embryo 5-3% O226-May-9831.2187538443

Embryo 5-3% O229-May-9863.3375565223

Embryo 5-3% O22-Jun-98105.531853742

Embryo 5-3% O26-Jun-98147.5521485042Sum of PD#totalDay# total

Embryo 5-3% O29-Jun-98179.1952350514Cell type036101114172124283438

Embryo 5-3% O213-Jun-982111.27361717Embryo 5-3% + UV01.21875384433.33755652235.5318537425.93239167167.004341513510.109796108311.936259853314.775547638616.9497409717

Embryo 5-3% O216-Jun-982413.0685528328Embryo 5-3% O201.21875384433.33755652235.5318537427.55214850429.195235051411.2736171713.068552832815.078138881717.795495431920.1393769679

Embryo 5-3% O220-Jun-982815.0781388817Embryo 6- 3% + UV01.32048708093.62985732255.84605347786.18633088267.326318599910.466006370912.135033136414.938260172817.3738887669

Embryo 5-3% O226-Jun-983417.7954954319Embryo 6- 3% O201.32048708093.62985732255.84605347787.99769585249.573008183111.320037460612.839411619714.547230868217.664925910920.4212375939

Embryo 5-3% O230-Jun-983820.1393769679

Embryo 5-3% + UV23-May-9800

Embryo 5-3% + UV26-May-9831.2187538443

Embryo 5-3% + UV29-May-9863.3375565223

Embryo 5-3% + UV2-Jun-98105.531853742

Embryo 5-3% + UV3-Jun-98115.9323916716

Embryo 5-3% + UV9-Jun-98177.0043415135

Embryo 5-3% + UV16-Jun-982410.1097961083

Embryo 5-3% + UV20-Jun-982811.9362598533

Embryo 5-3% + UV26-Jun-983414.7755476386

Embryo 5-3% + UV30-Jun-983816.9497409717

Embryo 6- 3% O223-May-9800

Embryo 6- 3% O226-May-9831.3204870809

Embryo 6- 3% O229-May-9863.6298573225

Embryo 6- 3% O22-Jun-98105.8460534778

Embryo 6- 3% O26-Jun-98147.9976958524

Embryo 6- 3% O29-Jun-98179.5730081831

Embryo 6- 3% O213-Jun-982111.3200374606

Embryo 6- 3% O216-Jun-982412.8394116197

Embryo 6- 3% O220-Jun-982814.5472308682

Embryo 6- 3% O226-Jun-983417.6649259109

Embryo 6- 3% O230-Jun-983820.4212375939

Embryo 6- 3% + UV23-May-9800

Embryo 6- 3% + UV26-May-9831.3204870809

Embryo 6- 3% + UV29-May-9863.6298573225

Embryo 6- 3% + UV2-Jun-98105.8460534778

Embryo 6- 3% + UV3-Jun-98116.1863308826

Embryo 6- 3% + UV9-Jun-98177.3263185999

Embryo 6- 3% + UV16-Jun-982410.4660063709

Embryo 6- 3% + UV20-Jun-982812.1350331364

Embryo 6- 3% + UV26-Jun-983414.9382601728

Embryo 6- 3% + UV30-Jun-983817.3738887669

Sheet3

0000

0000

0000

0000

0000

0000

0000

0000

0000

0000

0000

0000

Embryo 5-3% + UV

Embryo 5-3% O2

Embryo 6- 3% + UV

Embryo 6- 3% O2

days

Population doublings

Sheet2

Cell typeCumm cell#PassageDateDay# totalPD#totalCell typeCumm cell#PassageDateDay# totalPD#total

Embryo 5-20% O211910000123-May-9800Embryo 5-20% O211910000123-May-9800

Embryo 5-20% O227720000226-May-9831.2187538443Embryo 5-20% O227720000226-May-9831.2187538443Sum of PD#totalDay# totalSum of Cumm cell#Day# total

Embryo 5-20% O288703999.9999996329-May-9862.8968257494Embryo 5-20% O288703999.9999996329-May-9862.8968257494Cell type0361014172124283334384954Cell type0361014172124283334384954

Embryo 5-20% O2217029119.99999942-Jun-98104.1876433117Embryo 5-20% O2217029119.99999942-Jun-98104.1876433117Embryo 5-20% O201.21875384432.89682574944.18764331174.03028698385.15358147577.533017607110.84529553212.5112946048Embryo 5-20% O2119100002772000088703999.9999996217029119.999999194602777.599999423930214.3999972205850215.5946521911445474.906969532320307.0378

Embryo 5-20% O2194602777.59999956-Jun-98144.0302869838Embryo 5-20% O2194602777.59999956-Jun-98144.0302869838Embryo 5-3% O201.21875384433.33755652235.5318537427.55214850429.195235051411.2736171713.068552832815.078138881717.795495431920.1393769679Embryo 5-3% O21191000027720000120397200551017851.9999972235295752.946656981573735.036729485513074.305102314730367.838411987314281.1622709503236922.4413755244766109.6

Embryo 5-20% O2423930214.399997613-Jun-98215.1535814757Embryo 5-20% O2423930214.399997613-Jun-98215.1535814757Embryo 6- 3% O201.32048708093.62985732255.84605347787.99769585249.573008183111.320037460612.839411619714.547230868217.664925910920.4212375939Embryo 6- 3% O21202000030020000148799133.333333691419972.8888873072209412.869629155184050.3514430730901129.01388095249903.1704287777816350.3572497911445921.0916877555002940.2

Embryo 5-20% O22205850215.59465725-Jun-98337.5330176071Embryo 5-20% O22205850215.59465725-Jun-98337.5330176071Embryo 6-20% O201.32048708093.34078184314.96683153595.70379713017.0429345158.02860773359.0209082086Embryo 6-20% O21202000030020000121781133.333333375897764.888888626496274.8148131585035575.281483138746336.822216244079538.98765

Embryo 5-20% O221911445474.9069811-Jul-984910.845295532Embryo 5-20% O221911445474.9069811-Jul-984910.845295532



Embryo 5-3% O227720000226-May-9831.2187538443Embryo 5-3% O227720000226-May-9831.2187538443


Embryo 5-3% O2551017851.99999742-Jun-98105.531853742Embryo 5-3% O2551017851.99999742-Jun-98105.531853742








Embryo 5-3% + UV200000012-Jun-9805.531853742Embryo 6-20% O212020000123-May-9800

Embryo 5-3% + UV264000023-Jun-9815.9323916716Embryo 6-20% O230020000226-May-9831.3204870809

Embryo 5-3% + UV555000039-Jun-9877.0043415135Embryo 6-20% O2121781133.333333329-May-9863.3407818431

Embryo 5-3% + UV47767000.0000001416-Jun-981410.1097961083Embryo 6-20% O2375897764.88888842-Jun-98104.9668315359



Embryo 5-3% + UV5472131671.53123730-Jun-982816.9497409717Embryo 6-20% O23138746336.82221711-Jul-98498.0286077335

Embryo 6-20% O212020000123-May-9800Embryo 6-20% O26244079538.98765816-Jul-98549.0209082086

Embryo 6-20% O230020000226-May-9831.3204870809Embryo 6- 3% O212020000123-May-9800

Embryo 6-20% O2121781133.333333329-May-9863.3407818431Embryo 6- 3% O230020000226-May-9831.3204870809

Embryo 6-20% O2375897764.88888842-Jun-98104.9668315359Embryo 6- 3% O2148799133.333333329-May-9863.6298573225

Embryo 6-20% O2626496274.81481356-Jun-98145.7037971301Embryo 6- 3% O2691419972.88888742-Jun-98105.8460534778

Embryo 6-20% O21585035575.28148625-Jun-98337.042934515Embryo 6- 3% O23072209412.8696256-Jun-98147.9976958524

Embryo 6-20% O23138746336.82221711-Jul-98498.0286077335Embryo 6- 3% O29155184050.3514469-Jun-98179.5730081831

Embryo 6-20% O26244079538.98765816-Jul-98549.0209082086Embryo 6- 3% O230730901129.013713-Jun-982111.3200374606

Embryo 6- 3% O212020000123-May-9800Embryo 6- 3% O288095249903.1704816-Jun-982412.8394116197

Embryo 6- 3% O230020000226-May-9831.3204870809Embryo 6- 3% O2287777816350.357920-Jun-982814.5472308682

Embryo 6- 3% O2148799133.333333329-May-9863.6298573225Embryo 6- 3% O22497911445921.091026-Jun-983417.6649259109

Embryo 6- 3% O2691419972.88888742-Jun-98105.8460534778Embryo 6- 3% O216877555002940.21130-Jun-983820.4212375939

Embryo 6- 3% O23072209412.8696256-Jun-98147.9976958524

Embryo 6- 3% O29155184050.3514469-Jun-98179.5730081831

Embryo 6- 3% O230730901129.013713-Jun-982111.3200374606

Embryo 6- 3% O288095249903.1704816-Jun-982412.8394116197

Embryo 6- 3% O2287777816350.357920-Jun-982814.5472308682

Embryo 6- 3% O22497911445921.091026-Jun-983417.6649259109

Embryo 6- 3% O216877555002940.21130-Jun-983820.4212375939

Embryo 6- 3% + UV200000012-Jun-9805.8460534778

Embryo 6- 3% + UV253200023-Jun-9816.1863308826

Embryo 6- 3% + UV558000039-Jun-9877.3263185999

Embryo 6- 3% + UV49178400416-Jun-981410.4660063709

Embryo 6- 3% + UV156387312520-Jun-981812.1350331364

Embryo 6- 3% + UV1091583437.76626-Jun-982414.9382601728

Embryo 6- 3% + UV5905466398.28157730-Jun-982817.3738887669

Sheet2

0000

0000

0000

0000

0000

0000

0000

0000

0000

0000

0000

0000

0000

0000

Embryo 5-20% O2

Embryo 5-3% O2

Embryo 6- 3% O2

Embryo 6-20% O2

Days


Sheet1

0000

0000

0000

0000

0000

0000

0000

0000

0000

0000

0000

0000

0000

0000

Embryo 5-20% O2

Embryo 5-3% O2

Embryo 6- 3% O2

Embryo 6-20% O2

Days

Cummulative cell number

PDWorksheet

Embryo 5-20% O2DATA

Cumm cell#PassageDateDay# totalPD#total

11910000123-May-9800

27720000226-May-9831.2187538443

88703999.9999996329-May-9862.8968257494

217029119.99999942-Jun-98104.1876433117

194602777.59999956-Jun-98144.0302869838

423930214.399997613-Jun-98215.1535814757

2205850215.59465725-Jun-98337.5330176071

21911445474.9069811-Jul-984910.845295532

69532320307.0378916-Jul-985412.5112946048

Embryo 5-3% O2DATA


11910000123-May-9800

27720000226-May-9831.2187538443

120397200329-May-9863.3375565223

551017851.99999742-Jun-98105.531853742

2235295752.9466556-Jun-98147.5521485042

6981573735.036769-Jun-98179.1952350514

29485513074.305713-Jun-982111.27361717

102314730367.838816-Jun-982413.0685528328

411987314281.162920-Jun-982815.0781388817

2709503236922.441026-Jun-983417.7954954319

13755244766109.61130-Jun-983820.1393769679

Embryo 5-3% + UVDATA


200000012-Jun-9805.531853742

264000023-Jun-9815.9323916716

555000039-Jun-9877.0043415135

47767000.0000001416-Jun-981410.1097961083

169413626.666667520-Jun-981811.9362598533

1212436854.84444626-Jun-982414.7755476386

5472131671.53123730-Jun-982816.9497409717

Embryo 6-20% O2DATA


12020000123-May-9800

30020000226-May-9831.3204870809

121781133.333333329-May-9863.3407818431

375897764.88888842-Jun-98104.9668315359

626496274.81481356-Jun-98145.7037971301

1585035575.28148625-Jun-98337.042934515

3138746336.82221711-Jul-98498.0286077335

6244079538.98765816-Jul-98549.0209082086

Embryo 6- 3% O2DATA


12020000123-May-9800

30020000226-May-9831.3204870809

148799133.333333329-May-9863.6298573225

691419972.88888742-Jun-98105.8460534778

3072209412.8696256-Jun-98147.9976958524

9155184050.3514469-Jun-98179.5730081831

30730901129.013713-Jun-982111.3200374606

88095249903.1704816-Jun-982412.8394116197

287777816350.357920-Jun-982814.5472308682

2497911445921.091026-Jun-983417.6649259109

16877555002940.21130-Jun-983820.4212375939

Embryo 6- 3% + UVDATA


200000012-Jun-9805.8460534778

253200023-Jun-9816.1863308826

558000039-Jun-9877.3263185999

49178400416-Jun-981410.4660063709

156387312520-Jun-981812.1350331364

1091583437.76626-Jun-982414.9382601728

5905466398.28157730-Jun-982817.3738887669

isolations

Embryo 5-20% O2DATADATADATA

Cumm cell#PassageDateDay# totalDay#increm>Days/>PDCell# (out)PD#totalPD# incremCell# (in)# of T150 flasks# of pelletscell# per pelletLabel in the tubes

119100001 (thawed)23-May-9800000119100002 in 3% O2

27720000226-May-98332.4615306972277200001.21875384431.2187538443300000030Rest of cells frozen to repeat

88703999.9999998329-May-98631.787766061196000002.89682574941.678071905130000003166000005A-20%

217029119.99999942-Jun-981043.098811262573400004.18764331171.290817562330000003143400005B-20%

194602777.59999956-Jun-98144-25.420013623826900004.0302869838-0.1573563279269000030

423930214.399998613-Jun-982176.231669478258600005.15358147571.12329449199000003149600005C-20%(senesc)

2205850215.59466725-Jun-9833125.043211642146830007.53301760712.37943613156000003140830005D-20%

21911445474.9069811-Jul-9849164.8305125243596000010.8452955323.312277924915000003144600005E - 20%

69532320307.0379916-Jul-985453.0012021505476000012.51129460481.66599907281500003146100005F-20%

010-35938-359920000

011-3593800000

012-3593800000

013-3593800000



119100001 (thawed)23-May-9800000119100002 in 3% O2


120397200329-May-98631.4158940004130300003.33755652232.1188026781300000031100300005A-3%

55101785242-Jun-981041.8229071086137300005.5318537422.194297219730000003 and UV (1)177300005B-3%

2235295752.9466756-Jun-981441.9799091077121700007.55214850422.020294762230000003191700005C-3%

6981573735.0367569-Jun-981731.825832002193700009.19523505141.643086547230000003163700005D-3%

29485513074.3052713-Jun-982141.92457390971267000011.273617172.078382118630000003196700005E - 3%

102314730367.839816-Jun-982431.67136909821041000013.06855283281.794935662830000003174100005F-3%

411987314281.164920-Jun-982841.99045967821208000015.07813888172.009586048830000003190800005G-3%

2709503236922.451026-Jun-983462.20802823961973000017.79549543192.7173565502300000031167300005H-3%

13755244766109.61130-Jun-983841.70657089051523000020.13937696792.3438815364000000Frozen1112300005I-3%

Embryo 5-3% + UVDATADATADATA


20000001 (see 3% O2)2-Jun-980005.531853742020000001

2640000UV before passage 23-Jun-98112.496642455426400005.93239167160.4005379296264000030

555000039-Jun-98765.59727681855500007.00434151351.071949841915000003140500005A-UV

47767000416-Jun-981472.25409832481291000010.10979610833.105454594830000003199100005B-UV

169413626.666667520-Jun-981842.19002430841064000011.93625985331.82646374530000003176400005C-UV

1212436854.84444626-Jun-982462.11320600572147000014.77554763862.8392877854300000031184700005D-UV

5472131671.53125730-Jun-982841.83976279351354000016.94974097172.17419333314000000Frozen195400005E - UV



120200001 (thawed)23-May-9800000120200002 in 3% O2


121781133.333334329-May-98631.4849318308121700003.34078184312.020294762230000003191700006A-20%

375897764.88888942-Jun-981042.459949420992600004.96683153591.626049692830000003162600006B-20%

626496274.81481656-Jun-981445.427661795450000005.70379713010.73696559423000003147000006C-20%(senesc)

1585035575.28148625-Jun-98331914.18823805087590007.0429345151.339137384975900030

3138746336.82223711-Jul-98491616.232560345215030008.02860773350.9856732185150300030

6244079538.98766816-Jul-985455.038796337529900009.02090820860.99230047521500003128400006D-20%

09-35938-359920000

010-3593800000

011-3593800000

012-3593800000

013-3593800000

Embryo 6- 3% O2DATADATADATA


120200001 (thawed)23-May-9800000120200002 in 3% O2


148799133.333334329-May-98631.2990554507148700003.62985732252.3093702416300000031118700006A-3%

691419972.8888942-Jun-981041.8048943864139400005.84605347782.216196155430000003 and UV (1)179400006B-3%

3072209412.8696356-Jun-981441.8590450008133300007.99769585242.1516423746300000031103300006C-3%

9155184050.3515169-Jun-981731.904384255589400009.57300818311.575312330730000003159400006D-3%

30730901129.0132713-Jun-982142.2896010111007000011.32003746061.747029277530000003170700006E - 3%

88095249903.1712816-Jun-982431.9744971849860000012.83941161971.519374159130000003156000006F-3%

287777816350.359920-Jun-982842.3421682379980000014.54723086821.707819248530000003168000006G-3%

2497911445921.121026-Jun-983461.92449868182604000017.66492591093.1176950427300000031230400006H-3%

16877555002940.31130-Jun-983841.45121468842027000020.42123759392.7563116834000000Frozen1162700006I-3%

Embryo 6- 3% + UVDATADATADATA


20000001 (see 3% O2)2-Jun-980005.8460534778020000001


558000039-Jun-98765.263214602255800007.32631859991.139987717315000003140800006A-UV

49178400416-Jun-981472.22952105771322000010.46600637093.139687771300000031102200006B-UV

156387312520-Jun-981842.3966062634954000012.13503313641.669026765530000003165400006C-UV

1091583437.76626-Jun-982462.14039031522094000014.93826017282.8032270364300000031179400006D-UV

5905466398.28159730-Jun-982841.64228651681623000017.37388876692.43562859414000000Frozen1122300006E - UV

&F

Page &P

rescues6

MEF-Embryo 6

IDsamplecells(x106)lysis volumeTEOD260OD280260/280conc(ug/ul)DNA (ug)new TEnew concentugDNA/106 cellsdate isolated

0NT600ul07/22/02

16A-3%600ul1000.5590.3021.852.80279.52.807/22/02

26B-3%600ul1000.3060.1681.821.531531.537/22/02

36C-3%600ul1000.4510.241.882.26225.52.267/22/02

46D-3%600ul1000.4340.2341.852.172172.177/22/02

56E-3%600ul1000.2260.1261.791.131131.137/22/02

66F-3%600ul1000.320.1691.891.601601.607/22/02

76G-3%600ul1000.3380.1791.891.691691.697/22/02

86H-3%600ul1000.7080.3821.853.543543001.187/22/02

96I-3%600ul1000.8650.4521.914.33432.53001.447/22/02

106A-20%600ul1000.5460.2911.882.732732001.377/22/02

116B-20%600ul1000.4660.2541.832.332332001.177/22/02

126C-20%600ul1000.2210.1211.831.11110.51.117/22/02

136D-20%600ul1000.2510.1331.891.26125.51.267/22/02

146A-3%UV600ul1000.2020.1081.871.011011.017/22/02

156B-3%UV600ul1001.0080.681.485.045043001.687/22/02

166C-3%UV600ul1000.3750.2051.831.88187.51.887/22/02

176D-3%UV600ul1000.6070.3321.833.04303.52001.527/22/02

186E-3%UV600ul1000.3490.1861.881.75174.51.757/22/02

MEF-Embryo 5


0NT600ul0.3080.14908/16/02

15A-3%600ul1000.5230.2332.242.62261.58/16/02

25B-3%600ul1000.4010.1842.182.01200.58/16/02

35C-3%600ul1000.3940.1822.161.971978/16/02

45D-3%600ul1000.3470.1552.241.74173.58/16/02

55E-3%600ul1000.480.002.402408/16/02

65F-3%600ul1000.3470.162.171.74173.58/16/02

75G-3%600ul1000.4240.1892.242.122128/16/02

85H-3%600ul1000.6440.2892.233.223228/16/02

95I-3%600ul1000.440.1992.212.202208/16/02

105A-20%600ul1000.3430.1552.211.72171.58/16/02

115B-20%600ul1000.2440.112.221.221228/16/02

125C-20%600ul1000.2920.1412.071.461468/16/02

135D-20%600ul1000.2790.1252.231.40139.58/16/02

145E-20%600ul1000.310.142.211.551558/16/02

155F-20%600ul1000.2920.1342.181.46146

165A-3%UV600ul1000.2430.1122.171.22121.58/16/02

175B-3%UV600ul1000.520.2382.182.602608/16/02

185C-3%UV600ul1000.4220.1912.212.112118/16/02

195D-3%UV600ul1000.5770.2582.242.89288.58/16/02

205E-3%UV600ul1000.4960.2312.152.482488/16/02

MEF-Embryo 6


0NT600ul0.2630.121.3208/26/02

16E-20%600ul1000.2750.1282.151.38137.58/26/02

26F-20%600ul2000.5010.2282.202.515018/26/02

35G-20%600ul1000.3630.1612.251.82181.58/26/02

45H-20%600ul1000.7340.3332.203.673678/26/02

5Hum clone12F600ul2000.3440.1622.121.723448/26/02

AVERAGES6

Rescue 7-25-02Rescue 7-29-02Rescue 7-29-02Rescue 8-12-02

IDsampleconc(ug/ul)ug DNADNA ulH2OTCcellsX-galP-galMF(x10-5)Day# TotalPD# TotalHarvestnotesIDsampleconc(ug/ul)ug DNADNA ulH2OTCcellsX-galP-galMF(x10-5)Day# TotalPD# TotalnotesMFdaysPDIDsampleconc(ug/ul)ug DNADNA ulH2OTCcellsX-galP-galMF(x10-5)Day# TotalPD# TotalnotesMFdaysPDIDsampleconc(ug/ul)ug DNADNA ulH2OTCcellsX-galP-galMF(x10-5)Day# TotalPD# Total

0NT004.57/12/020100NT004.57/12/02000NT004.57/12/020100NT004.57/12/02000

16A-3%2.80207.1650.844.57/12/02450398.6763.62985732258.673.6298573225616A-3%2.80207.1650.844.57/12/02216146.4863.62985732256.4863.629857322516A-3%2.80207.1650.844.57/12/021470.0063.62985732256.4863.629857322516A-3%2.80207.1650.844.57/12/02150128.0063.6298573225

26B-3%1.532013.0744.934.57/12/023463710.69105.846053477810.695.84605347781026B-3%1.532013.0744.934.57/12/02157138.28105.84605347788.28105.846053477826B-3%1.532013.0744.934.57/12/021531711.11105.84605347788.28105.846053477826D-3%2.17209.2248.784.57/12/021302317.69179.5730081831

36C-3%2.26208.8749.134.57/12/023465415.61147.997695852415.617.99769585241436C-3%2.26208.8749.134.57/12/022213114.03147.997695852414.03147.997695852436C-3%2.26208.8749.134.57/12/021281814.06147.997695852414.03147.997695852436E-3%1.132017.7040.304.57/12/026327311.552111.3200374606

46D-3%2.17209.2248.784.57/12/02672943.28179.573008183119.166666666711.32003746062146D-3%2.17209.2248.784.57/12/021642716.46179.573008183116.46179.573008183146D-3%2.17209.2248.784.57/12/02811619.75179.573008183116.46179.573008183146G-3%1.692011.8346.174.57/12/021452114.482814.5472308682

56E-3%1.132017.7040.304.57/12/023606919.172111.320037460618.965517241412.83941161972456E-3%1.132017.7040.304.57/12/02266228.272111.32003746068.272111.320037460656E-3%1.132017.7040.304.57/12/022433213.172111.32003746068.272111.320037460656H-3%1.182016.9541.054.57/12/023483911.213417.6649259109

66F-3%1.602012.5045.504.57/12/022324418.972412.839411619710.112359550614.54723086822866F-3%1.602012.5045.504.57/12/021332518.802412.839411619718.802412.839411619766F-3%1.602012.5045.504.57/12/02761418.422412.839411619718.802412.839411619766A-20%1.372014.6543.354.57/12/02218146.4263.3407818431

76G-3%1.692011.8346.174.57/12/021781810.112814.547230868214.095744680917.66492591093476G-3%1.692011.8346.174.57/12/02431330.232814.54723086829.523417.664925910976G-3%1.692011.8346.174.57/12/02751114.672814.54723086829.523417.664925910976B-20%1.172017.1740.834.57/12/0212197.44104.9668315359

86H-3%1.182016.9541.054.57/12/023765314.103417.664925910911.006289308220.42123759393886H-3%1.182016.9541.054.57/12/02210209.523417.664925910910.233820.421237593986H-3%1.182016.9541.054.57/12/021701810.593417.664925910910.233820.421237593986C-20%1.112018.1039.904.57/12/022033014.78145.7037971301

96I-3%1.442013.8744.134.57/12/023183511.013820.421237593996I-3%1.442013.8744.134.57/12/022152210.233820.421237593996I-3%1.442013.8744.134.57/12/02149149.403820.421237593996D-20%1.262015.9442.064.57/12/021022827.45549.0209082086

106A-20%1.372014.6543.354.57/12/021832212.0263.3407818431106A-20%1.372014.6543.354.57/12/0210798.4163.34078184318.411214953363.3407818431106A-20%1.372014.6543.354.57/12/02150138.6763.34078184318.411214953363.3407818431106A-3%UV1.012019.8038.204.57/12/02152213140.1377.3263185999

116B-20%1.172017.1740.834.57/12/022963311.15104.9668315359116B-20%1.172017.1740.834.57/12/021151815.65104.966831535915.652173913104.9668315359116B-20%1.172017.1740.834.57/12/0223521.74104.966831535915.652173913104.9668315359116E-3%UV1.752011.4646.544.57/12/02298353118.462817.3738887669

126C-20%1.112018.1039.904.57/12/023415114.96145.7037971301senescent126C-20%1.112018.1039.904.47/12/029444.44145.7037971301senescent15.3333333333549.0209082086126C-20%1.112018.1039.904.47/12/02902022.22145.7037971301senescent15.3333333333549.020908208612#30 ctrl1.962010.204081632747.79591836734.57/12/02720385.28

136D-20%1.262015.9442.064.57/12/021804625.56549.0209082086136D-20%1.262015.9442.064.57/12/021502315.33549.0209082086136D-20%1.262015.9442.064.57/12/02872528.74549.0209082086

146A-3%UV1.012019.8038.204.57/12/02247388157.0977.3263185999146A-3%UV1.012019.8038.204.57/12/02146198135.6277.3263185999146A-3%UV1.012019.8038.204.57/12/02701825.7177.3263185999

156B-3%UV1.682011.9046.104.57/12/02320373116.561410.4660063709156B-3%UV1.682011.9046.104.57/12/02133134100.751410.4660063709156B-3%UV1.682011.9046.104.57/12/02168173102.981410.4660063709

166C-3%UV1.882010.6747.334.57/12/0240640499.511812.1350331364166C-3%UV1.882010.6747.334.57/12/0212411794.351812.1350331364166C-3%UV1.882010.6747.334.57/12/021049389.421812.1350331364

176D-3%UV1.522013.1844.824.57/12/0224422692.622414.9382601728176D-3%UV1.522013.1844.824.57/12/0230625884.312414.9382601728176D-3%UV1.522013.1844.824.57/12/02390391100.262414.9382601728

186E-3%UV1.752011.4646.544.57/12/02380428112.632817.3738887669186E-3%UV1.752011.4646.544.57/12/02231303131.172817.3738887669186E-3%UV1.752011.4646.544.57/12/02288490170.142817.3738887669

19#30 ctrl1.962010.2047.804.57/12/02632507.9119#30 ctrl1.962010.204081632747.79591836734.57/12/022532610.2819#30 ctrl1.962010.2047.804.57/12/02261186.90

206D-3%2.17209.2248.784.57/12/02421126.19

216E-3%1.132017.7040.304.57/12/02111109.01

226G-3%1.692011.8346.174.57/12/02321443.75

236H-3%1.182016.9541.054.57/12/021332216.54

246C-20%1.112018.1039.904.47/12/02361130.56

256D-20%1.262015.9442.064.57/12/02491428.57

RESCUE 8-29-02

IDsampleconc(ug/ul)ug DNADNA ulH2OTCcellsX-galP-galMF(x10-5)

0NT1.322015.242.84.58/27/02000

16A-3%2.80207.250.84.58/27/022312510.82

26A-3%2.80207.250.84.58/27/021692615.38

36I-3%1.442013.944.14.58/27/021813217.68

46I-3%1.442013.944.14.58/27/021392417.27

56A-20%1.372014.743.34.58/27/02141117.80ctrl

66A-20%1.372014.743.34.58/27/02220198.64ctrl

76E-20%1.382014.543.54.58/27/021584830.38

86F-20%2.51208.050.04.58/27/021444631.94

9#30 ctrl1.962010.247.84.58/27/0233233611.15

1012F1.722011.646.44.58/27/02000.00

AVERAGES6

00

00

00

00

0

0

0

0

0

6-3%O2

6-20%O2

PD#Total

mutation frequency (x10-5)

MEF 6

RESCUES 5

00

00

00

00

0

0

0

0

6-3%02

6-20%

PD#Total

Mutation Frequency (x10-5)

MEF6* #1

Averages5

00

00

00

00

00

0

0

0

6-3%O2

6-3%02UV

PD#Total


MEF6-UV#1

combined growth curves

00

00

00

00

0

0

0

0

6-3%02

6-20%

Day#Total


MEF6* #1

00

00

00

00

00

0

0

0

6-3%O2

6-3%02UV

Day#Total


MEF6-UV#1

00

00

00

0

0

0

0

0

6-3%02

6-20%

PD#Total


MEF6* #2

00

00

00

00

00

0

0

0

6-3%O2

6-3%02UV

PD#Total


MEF6-UV#2

00

00

00

0

0

0

0

0

6-3%02

6-20%

Days


MEF6* #2

00

00

00

00

00

0

0

0

6-3%O2

6-3%02UV

Days


MEF6-UV#2

Averages

IDsample7/24/987/28/988/5/028/5/028/12/028/29/029/23/029/23/02AveragestdevPD

0NT00

16A-3%8.676.4812.248.0010.829.22.33.6

26B-3%10.698.2811.119.71.55.8

36C-3%15.6114.0314.0614.00.98.0

46D-3%16.4619.7517.6918.01.79.6

56E-3%19.178.2713.179.0111.5510.54.311.3

66F-3%18.9718.8018.4218.60.312.8

76G-3%10.1114.6714.4814.62.614.5

86H-3%14.109.5210.5916.5411.2112.02.917.7

96I-3%11.0110.239.4017.6817.2713.14.020.4

106A-20%12.028.418.676.427.808.648.71.83.340782

116B-20%11.1515.657.4411.54.14.966832

126C-20%14.9622.2214.7818.54.25.703797

136D-20%25.5615.3328.7428.5727.4525.05.69.0209082

6E-20%30.3840.0031.1133.85.413.8868270

6F-20%31.9434.8333.6033.51.416.9537773

146A-3%UV157.09135.62140.13137.911.3

156B-3%UV116.56100.75102.98101.98.6

166C-3%UV99.5194.3589.4291.95.0

176D-3%UV92.6284.31100.2692.38.0

186E-3%UV112.63131.17118.46124.89.5

19#30 ctrl7.9110.286.905.287.52.1

Cumulative7/24/987/28/98SUMMF(x10-5)3%O2

IDsampleX-galP-galX-galP-galX-galP-galX-galP-gal20%O2

0NT010010

16A-3%4503921614666537.96

26B-3%3463715713503509.94

36C-3%34654221315678514.99

46D-3%6729164272315624.24

56E-3%36069266226269114.54

66F-3%23244133253656918.90

76G-3%178181781810.11

86H-3%37653210205867312.46

96I-3%31835215225335710.69

106A-20%1832210792903110.69

116B-20%29633115184115112.41

126C-20%341513415114.96

136D-20%18046150233306920.91

146A-3%UV247388146198393586149.11

156B-3%UV320373133134453507111.92

166C-3%UV40640412411753052198.30

176D-3%UV24422630625855048488.00

186E-3%UV380428231303611731119.64

19#30 ctrl6325025326885768.59

002.29180274962.29180274961.84988161521.8498816152

001.52820506961.52820506964.11345175544.1134517554

000.90205882840.90205882844.24737156894.2473715689

001.66227392291.66227392295.62109280885.6210928088

004.34485506164.34485506165.35562804185.3556280418

000.27873914830.27873914831.44869034531.4486903453

02.57798123572.5779812357

02.87247411682.8724741168

04.02038027144.0203802714

3%O2

20%O2

PD


Average MF for MEF 6 (PD)

002.29180274962.29180274961.84988161521.8498816152

001.52820506961.52820506964.11345175544.1134517554

000.90205882840.90205882844.24737156894.2473715689

001.66227392291.66227392295.62109280885.6210928088

04.34485506164.3448550616

00.27873914830.2787391483

02.57798123572.5779812357

02.87247411682.8724741168

04.02038027144.0203802714

MEF6-3%O2

MEF6-20%O2

Days Total


Average MF for MEF6 (vs Days)

002.29180274962.291802749611.318905089511.3189050895

001.52820506961.52820506968.55872925938.5587292593

000.90205882840.90205882845.04255893765.0425589376

001.66227392291.66227392297.97372901167.9737290116

004.34485506164.34485506169.47949101369.4794910136

00.27873914830.2787391483

02.57798123572.5779812357

02.87247411682.8724741168

04.02038027144.0203802714

MEF6-3%O2

MEF6-3%O2UV

PD


Average for MEF6-UV

002.29180274962.291802749611.318905089511.3189050895

001.52820506961.52820506968.55872925938.5587292593

000.90205882840.90205882845.04255893765.0425589376

001.66227392291.66227392297.97372901167.9737290116

004.34485506164.34485506169.47949101369.4794910136

00.27873914830.2787391483

02.57798123572.5779812357

02.87247411682.8724741168

04.02038027144.0203802714

MEF6-3%O2

MEF6-3%O2UV

Days


Average for MEF6-UV

Rescue 9-3-02Rescue 9-12-02Rescue 9-17-02Rescue 9-23-02

IDsampleconc(ug/ul)ug DNADNA ulH2OTCcellsX-galP-galMF(x10-5)IDsampleconc(ug/ul)ug DNADNA ulH2OTCcellsX-galP-galMF(x10-5)IDsampleconc(ug/ul)ug DNADNA ulH2OTCcellsX-galP-galMF(x10-5)IDsampleconc(ug/ul)ug DNADNA ulH2OTCcellsX-galP-galMF(x10-5)

0NT1.322015.1542.854.57/12/020100NT1.322015.1542.854.59/19/010200NT1.322015.1542.854.59/19/010100NT1.322015.1542.854.59/19/01000

15A-3%2.62207.6550.354.57/12/0210298.8215A-3%2.62207.6550.354.59/19/013083812.3415A-3%2.62207.6550.354.59/19/012352611.0615A-3%2.62207.6550.354.59/19/01313268.31

25B-3%2.01209.9848.024.57/12/02741013.5125B-3%2.01209.9848.024.59/19/01133107.5225B-3%2.01209.9848.024.59/19/01266145.2625B-3%2.01209.9848.024.59/19/0114296.34

35C-3%1.972010.1547.853.97/12/029499.5735C-3%1.972010.1547.854.59/19/011542616.8835C-3%1.972010.1547.854.59/19/012883311.4635C-3%1.972010.1547.854.59/19/01358236.42

45D-3%1.742011.5346.474.57/12/028344.8245D-3%1.742011.5346.474.59/19/01139117.9145D-3%1.742011.5346.474.09/19/01350349.7145D-3%1.742011.5346.474.59/19/013294714.29

55E-3%2.40208.3349.674.57/12/026669.0955E-3%2.40208.3349.674.59/19/011741810.3455E-3%2.40208.3349.674.59/19/013043411.1855E-3%2.40208.3349.674.59/19/012553011.76

65F-3%1.742011.5346.474.57/12/021061917.9265F-3%1.742011.5346.474.59/19/012402410.0065F-3%1.742011.5346.474.59/19/01376359.3165F-3%1.742011.5346.474.59/19/01324288.64

75G-3%2.12209.4348.574.57/12/029244.3575G-3%2.12209.4348.574.59/19/01160159.3875G-3%2.12209.4348.574.59/19/013203611.2575G-3%2.12209.4348.574.59/19/01402379.20

85H-3%3.22206.2151.794.47/12/028256.1085H-3%3.22206.2151.794.09/19/01219219.5985H-3%3.22206.2151.794.59/19/01306206.5485H-3%3.22206.2151.794.59/19/01431337.66

95I-3%2.20209.0948.914.57/12/02701217.1495I-3%2.20209.0948.914.59/19/01234114.7095I-3%2.20209.0948.914.39/19/0112564.8095I-3%2.20209.0948.914.59/19/01367359.54

105A-20%2.21209.0448.964.57/12/025347.55105A-20%2.21209.0448.964.59/19/01186158.06105A-20%2.21209.0448.964.59/19/01113108.85105A-20%2.21209.0448.964.59/19/01147106.80

115B-20%2.22209.0248.984.57/12/0218211.11115B-20%2.22209.0248.984.59/19/016369.52115B-20%2.22209.0248.984.59/19/0112597.20115B-20%2.22209.0248.984.59/19/012292711.79

125C-20%2.07209.6648.344.57/12/026369.52125C-20%2.07209.6648.344.59/19/012063617.48125C-20%2.07209.6648.344.59/19/011612314.29125C-20%2.07209.6648.344.59/19/01189168.47

135D-20%2.23208.9649.044.57/12/0238513.16135D-20%2.23208.9649.044.59/19/011271814.17135D-20%2.23208.9649.044.59/19/011391510.79135D-20%2.23208.9649.044.59/19/011542516.23

145E-20%2.21209.0348.974.57/12/0237718.92145E-20%2.21209.0348.974.59/19/011292620.16145E-20%2.21209.0348.974.59/19/01134139.70145E-20%2.21209.0348.974.59/19/011861910.22

155F-20%2.18209.1848.824.57/12/02841214.29155F-20%2.18209.1848.824.59/19/011071816.82155F-20%2.18209.1848.824.59/19/011262620.63155F-20%2.18209.1848.824.59/19/011482818.92

15a5G-20%1.822010.9947.014.57/12/022063717.9615a5G-20%1.822010.9947.014.59/19/011514026.4915a5G-20%1.822010.9947.014.59/19/0141010325.1215a5G-20%1.822010.9947.014.59/19/0129712241.08

15b5H-20%3.67205.4552.554.57/12/021815630.9415b5H-20%3.67205.4552.554.59/19/0165218027.6115b5H-20%3.67205.4552.554.59/19/0172018525.6915b5H-20%3.67205.4552.554.59/19/0140213333.08

165A-3%UV2.17209.2248.784.57/12/021538253.33165A-3%UV2.17209.2248.784.59/19/01140146104.29165A-3%UV2.17209.2248.784.59/19/01168208123.81165A-3%UV2.17209.2248.784.59/19/01172192111.63

175B-3%UV2.18209.1548.854.57/12/028891103.41175B-3%UV2.18209.1548.854.59/19/01196208106.12175B-3%UV2.18209.1548.854.59/19/01243280115.23175B-3%UV2.18209.1548.854.59/19/01315354112.38

185C-3%UV2.21209.0548.954.57/12/021127466.07185C-3%UV2.21209.0548.954.49/19/0183111133.73185C-3%UV2.21209.0548.954.49/19/01244266109.02185C-3%UV2.21209.0548.954.49/19/01321341106.23

195D-3%UV2.24208.9449.064.57/12/0225424797.24195D-3%UV2.24208.9449.064.59/19/0129728595.96195D-3%UV2.24208.9449.064.59/19/0147645896.22195D-3%UV2.24208.9449.064.59/19/0167666998.96

205E-3%UV2.15209.3148.694.57/12/0214610269.86205E-3%UV2.15209.3148.694.59/19/0152237772.22205E-3%UV2.15209.3148.694.59/19/0127323786.81205E-3%UV2.15209.3148.694.59/19/0141235084.95

21#30 ctrl1.962010.2047.804.57/12/02341288.2121#30 ctrl1.962010.2047.804.59/19/01736658.8321#30 ctrl1.962010.2047.804.59/19/01952626.5121#30 ctrl1.962010.2047.804.59/19/01632497.75

IDsample9/12/029/17/029/23/02AveragesstdevPD total

0NT00

15A-3%12.3411.068.3110.572.13.3375565223

25B-3%7.525.266.346.371.15.531853742

35C-3%16.8811.466.4211.595.27.5521485042

45D-3%7.919.7114.2910.643.39.1952350514

55E-3%10.3411.1811.7611.100.711.27361717

65F-3%10.009.318.649.320.713.0685528328

75G-3%9.3811.259.209.941.115.0781388817

85H-3%9.596.547.667.931.517.7954954319

95I-3%4.704.809.546.352.820.1393769679

105A-20%8.068.856.807.911.02.8968257494

115B-20%9.527.2011.799.502.34.1876433117

125C-20%17.4814.298.4713.414.65.1535814757

135D-20%14.1710.7916.2313.732.77.5330176071

145E-20%20.169.7010.2213.365.910.845295532

155F-20%16.8220.6318.9218.791.912.5112946048

15a5G-20%26.4925.1241.0830.908.818.5022494652

15b5H-20%27.6125.6933.0828.803.822.2012087368

165A-3%UV104.29123.81111.63113.249.97.0043415135

175B-3%UV106.12115.23112.38111.244.710.1097961083

185C-3%UV109.02106.23107.622.011.9362598533

195D-3%UV95.9696.2298.9697.051.714.7755476386

205E-3%UV72.2286.8184.9581.337.916.9497409717

21#30 ctrl8.836.517.757.701.2

002.06045891382.06045891381.03263062931.0326306293

001.12823378641.12823378642.29525595642.2952559564

005.23048614315.23048614314.56858980924.5685898092

003.28489319463.28489319462.74780256512.7478025651

000.71386190940.71386190945.89270276815.8927027681

000.6790505560.6790505561.90940713411.9094071341

001.13512629541.13512629548.84346180678.8434618067

001.54442810371.54442810373.83564857893.8356485789

02.76384933352.7638493335

MEF5-3%

MEF5-20%

PD Total

MF

009.86136164119.86136164112.06045891382.0604589138

004.65735737414.65735737411.12823378641.1282337864

001.96990321811.96990321815.23048614315.2304861431

001.6651838871.6651838873.28489319463.2848931946

007.94140718687.94140718680.71386190940.7138619094

00.6790505560.679050556

01.13512629541.1351262954

01.54442810371.5444281037

02.76384933352.7638493335

MEF5-3%

MEF5-3%UV

PD Total

MF

002.06045891382.06045891381.03263062931.0326306293

001.12823378641.12823378642.29525595642.2952559564

005.23048614315.23048614314.56858980924.5685898092

003.28489319463.28489319462.74780256512.7478025651

000.71386190940.71386190945.89270276815.8927027681

000.6790505560.6790505561.90940713411.9094071341

001.13512629541.13512629548.84346180678.8434618067

001.54442810371.54442810373.83564857893.8356485789

02.76384933352.7638493335

3%

20%

Population doublings total

Mutant frequency (x10-5)

00000000000000

3.33504167182.92700552112.69209835083.04073852143.10772421063.16581323783.6112035853.28066858832.84172474942.61967023531.21875384431.21875384431.32048708091.3204870809

6.29481182714.63040158256.1026203697.10193867786.50643940467.04617225074.79484889045.46494888274.68168433694.67538649882.89682574943.33755652233.34078184313.6298573225

8.59249237575.86367612159.416833498710.41421660268.98325310149.75986806556.43698921258.08475313276.72782611856.73428018794.18764331175.5318537424.96683153595.8460534778

11.12356186848.54974166512.536742962514.12864530810.670313789812.6330250468.473785568510.91730563488.47126927689.24186342874.03028698387.55214850425.70379713017.9976958524

13.291347959110.804738751715.834423511117.418479773215.59473043815.55268293778.756719531813.1712949019.783547201611.53758645325.15358147579.19523505147.0429345159.5730081831

15.679078112410.875128079620.813229288820.587335715720.991620590518.22774609297.533017607111.273617178.028607733511.3200374606

20.31521933712.87855898525.888679478423.097288529810.84529553213.06855283289.020908208612.8394116197

25.578253742915.954661964112.511294604815.078138881714.5472308682

19.320167877617.795495431917.6649259109

23.052075181920.139376967920.4212375939

28.3954830042

1-20%

1-3%

3-20%

3-3%

4-20%

4-3%

1-20% B

1-3% B

2-20%

2-3%

5-20%

5-3%

6-20%

6-3%

Days

PD

Chart8

9.2431113868.6599233282.29180274962.29180274961.84988161521.8498816152

9.695682944111.5450952211.52820506961.52820506964.11345175544.1134517554

14.044824660618.50027367270.90205882840.90205882844.24737156894.2473715689

17.969602915425.02284362021.66227392291.66227392295.62109280885.6210928088

10.49976072333.83028598224.34485506164.34485506165.35562804185.3556280418

18.609022556433.45863503950.27873914830.27873914831.44869034531.4486903453

14.57471264372.57798123572.5779812357

11.96507368832.87247411682.8724741168

13.11611313274.02038027144.0203802714

3%O2

20%O2


Mutant Frequency (x10-5)

Sheet3



Embryo 5-3% O226-May-9831.2187538443

Embryo 5-3% O229-May-9863.3375565223

Embryo 5-3% O22-Jun-98105.531853742







Embryo 5-3% O230-Jun-983820.1393769679


Embryo 5-3% + UV26-May-9831.2187538443

Embryo 5-3% + UV29-May-9863.3375565223

Embryo 5-3% + UV2-Jun-98105.531853742

Embryo 5-3% + UV3-Jun-98115.9323916716

Embryo 5-3% + UV9-Jun-98177.0043415135

Embryo 5-3% + UV16-Jun-982410.1097961083

Embryo 5-3% + UV20-Jun-982811.9362598533

Embryo 5-3% + UV26-Jun-983414.7755476386

Embryo 5-3% + UV30-Jun-983816.9497409717

Embryo 6- 3% O223-May-9800

Embryo 6- 3% O226-May-9831.3204870809

Embryo 6- 3% O229-May-9863.6298573225

Embryo 6- 3% O22-Jun-98105.8460534778

Embryo 6- 3% O26-Jun-98147.9976958524

Embryo 6- 3% O29-Jun-98179.5730081831

Embryo 6- 3% O213-Jun-982111.3200374606

Embryo 6- 3% O216-Jun-982412.8394116197

Embryo 6- 3% O220-Jun-982814.5472308682

Embryo 6- 3% O226-Jun-983417.6649259109

Embryo 6- 3% O230-Jun-983820.4212375939

Embryo 6- 3% + UV23-May-9800

Embryo 6- 3% + UV26-May-9831.3204870809

Embryo 6- 3% + UV29-May-9863.6298573225

Embryo 6- 3% + UV2-Jun-98105.8460534778

Embryo 6- 3% + UV3-Jun-98116.1863308826

Embryo 6- 3% + UV9-Jun-98177.3263185999

Embryo 6- 3% + UV16-Jun-982410.4660063709

Embryo 6- 3% + UV20-Jun-982812.1350331364

Embryo 6- 3% + UV26-Jun-983414.9382601728

Embryo 6- 3% + UV30-Jun-983817.3738887669

Sheet3

0000

0000

0000

0000

0000

0000

0000

0000

0000

0000

0000

0000

Embryo 5-3% + UV

Embryo 5-3% O2

Embryo 6- 3% + UV

Embryo 6- 3% O2

days


Sheet2

Cell typeCumm cell#PassageDateDay# totalPD#totalCell typeCumm cell#PassageDateDay# totalPD#totalSum of PD#totalDay# total

Embryo 5-20% O211910000123-May-9800Embryo 5-20% O211910000123-May-9800Cell type03610141721242833343849547784

Embryo 5-20% O227720000226-May-9831.2187538443Embryo 5-20% O227720000226-May-9831.2187538443Embryo 5-20% O201.21875384432.89682574944.18764331174.03028698385.15358147577.533017607110.84529553212.511294604818.502249465222.2012087368

Embryo 5-20% O288703999.9999996329-May-9862.8968257494Embryo 5-20% O288703999.9999996329-May-9862.8968257494Embryo 5-3% O201.21875384433.33755652235.5318537427.55214850429.195235051411.2736171713.068552832815.078138881717.795495431920.1393769679

Embryo 5-20% O2217029119.99999942-Jun-98104.1876433117Embryo 5-20% O2217029119.99999942-Jun-98104.1876433117Embryo 6- 3% O201.32048708093.62985732255.84605347787.99769585249.573008183111.320037460612.839411619714.547230868217.664925910920.4212375939

Embryo 5-20% O2194602777.59999956-Jun-98144.0302869838Embryo 5-20% O2194602777.59999956-Jun-98144.0302869838Embryo 6-20% O201.32048708093.34078184314.96683153595.70379713017.0429345158.02860773359.020908208613.886827023216.9537772671





Embryo 5-20% O24422255571527.6108-Aug-987718.5022494652Embryo 5-20% O24422255571527.6108-Aug-987718.5022494652MEF 5

Embryo 5-20% O257430359022238.51115-Aug-988422.2012087368Embryo 5-20% O257430359022238.51115-Aug-988422.2012087368




















Embryo 6-20% O230020000226-May-9831.3204870809Embryo 6-20% O2182077359356.8898-Aug-987713.8868270232

Embryo 6-20% O2121781133.333333329-May-9863.3407818431Embryo 6-20% O21525808271410.651015-Aug-988416.9537772671

Embryo 6-20% O2375897764.88888842-Jun-98104.9668315359Embryo 6-3% O212020000123-May-9800

Embryo 6-20% O2626496274.81481356-Jun-98145.7037971301Embryo 6-3% O230020000226-May-9831.3204870809

Embryo 6-20% O21585035575.28148625-Jun-98337.042934515Embryo 6-3% O2148799133.333333329-May-9863.6298573225

Embryo 6-20% O23138746336.82221711-Jul-98498.0286077335Embryo 6-3% O2691419972.88888742-Jun-98105.8460534778


Embryo 6-20% O2182077359356.8898-Aug-987713.8868270232Embryo 6-3% O29155184050.3514469-Jun-98179.5730081831


Embryo 6- 3% O212020000123-May-9800Embryo 6-3% O288095249903.1704816-Jun-982412.8394116197Sum of Cumm cell#Day# total

Embryo 6- 3% O230020000226-May-9831.3204870809Embryo 6-3% O2287777816350.357920-Jun-982814.5472308682Cell type03610141721242833343849547784

Embryo 6- 3% O2148799133.333333329-May-9863.6298573225Embryo 6-3% O22497911445921.091026-Jun-983417.6649259109Embryo 5-20% O2119100002772000088703999.9999996217029119.999999194602777.599999423930214.3999972205850215.5946521911445474.906969532320307.03784422255571527.657430359022238.5

Embryo 6- 3% O2691419972.88888742-Jun-98105.8460534778Embryo 6-3% O216877555002940.21130-Jun-983820.4212375939Embryo 5-3% O21191000027720000120397200551017851.9999972235295752.946656981573735.036729485513074.305102314730367.838411987314281.1622709503236922.4413755244766109.6

Embryo 6- 3% O23072209412.8696256-Jun-98147.9976958524Embryo 6-20% O21202000030020000121781133.333333375897764.888888626496274.8148131585035575.281483138746336.822216244079538.98765182077359356.881525808271410.65

Embryo 6- 3% O29155184050.3514469-Jun-98179.5730081831Embryo 6-3% O21202000030020000148799133.333333691419972.8888873072209412.869629155184050.3514430730901129.01388095249903.1704287777816350.3572497911445921.0916877555002940.2

Embryo 6- 3% O230730901129.013713-Jun-982111.3200374606

Embryo 6- 3% O288095249903.1704816-Jun-982412.8394116197

Embryo 6- 3% O2287777816350.357920-Jun-982814.5472308682

Embryo 6- 3% O22497911445921.091026-Jun-983417.6649259109

Embryo 6- 3% O216877555002940.21130-Jun-983820.4212375939

Embryo 6- 3% + UV200000012-Jun-9805.8460534778

Embryo 6- 3% + UV253200023-Jun-9816.1863308826

Embryo 6- 3% + UV558000039-Jun-9877.3263185999

Embryo 6- 3% + UV49178400416-Jun-981410.4660063709

Embryo 6- 3% + UV156387312520-Jun-981812.1350331364

Embryo 6- 3% + UV1091583437.76626-Jun-982414.9382601728

Embryo 6- 3% + UV5905466398.28157730-Jun-982817.3738887669

MEF 6

bold 3%

open 20%

Sheet2

0000

0000

0000

0000

0000

0000

0000

0000

0000

0000

0000

0000

0000

0000

0000

0000

Embryo 5-20% O2

Embryo 5-3% O2

Embryo 6- 3% O2

Embryo 6-20% O2

Days


Sheet1

0000

0000

0000

0000

0000

0000

0000

0000

0000

0000

0000

0000

0000

0000

0000

0000

Embryo 5-20% O2

Embryo 5-3% O2

Embryo 6-20% O2

Embryo 6-3% O2

Days

Cummulative Cell Number

PDWorksheet

00

00

00

00

00

00

00

00

00

00

00

20%

3%

Days


00

00

00

00

00

00

00

00

00

00

0

Embryo 6- 3% O2

Embryo 6-20% O2

Days


9.2431113868.6599233282.29180274962.29180274961.84988161521.8498816152

9.695682944111.5450952211.52820506961.52820506964.11345175544.1134517554

14.044824660618.50027367270.90205882840.90205882844.24737156894.2473715689

17.969602915425.02284362021.66227392291.66227392295.62109280885.6210928088

10.49976072333.83028598224.34485506164.34485506165.35562804185.3556280418

18.609022556433.45863503950.27873914830.27873914831.44869034531.4486903453

14.57471264372.57798123572.5779812357

11.96507368832.87247411682.8724741168

13.11611313274.02038027144.0203802714

3%O2

20%O2



9.2431113868.6599233282.29180274962.29180274961.84988161521.8498816152

9.695682944111.5450952211.52820506961.52820506964.11345175544.1134517554

14.044824660618.50027367270.90205882840.90205882844.24737156894.2473715689

17.969602915425.02284362021.66227392291.66227392295.62109280885.6210928088

10.49976072333.83028598224.34485506164.34485506165.35562804185.3556280418

18.609022556433.45863503950.27873914830.27873914831.44869034531.4486903453

14.57471264372.57798123572.5779812357

11.96507368832.87247411682.8724741168

13.11611313274.02038027144.0203802714

3%O2

20%O2



10.56940046347.90559824652.06045891382.06045891381.03263062931.0326306293

6.37332768549.5047341791.12823378641.12823378642.29525595642.2952559564

11.58867707413.40901696895.23048614315.23048614314.56858980924.5685898092

10.637889688213.73278716223.28489319463.28489319462.74780256512.7478025651

11.09791466513.35719502010.71386190940.71386190945.89270276815.8927027681

9.31682864918.79208982010.6790505560.6790505561.90940713411.9094071341

9.942993366530.8964861741.13512629541.13512629548.84346180678.8434618067

7.927200445828.7954611741.54442810371.54442810373.83564857893.8356485789

6.3458798142.76384933352.7638493335

3%

20%



10.56940046347.90559824652.06045891382.06045891381.03263062931.0326306293

6.37332768549.5047341791.12823378641.12823378642.29525595642.2952559564

11.58867707413.40901696895.23048614315.23048614314.56858980924.5685898092

10.637889688213.73278716223.28489319463.28489319462.74780256512.7478025651

11.09791466513.35719502010.71386190940.71386190945.89270276815.8927027681

9.31682864918.79208982010.6790505560.6790505561.90940713411.9094071341

9.942993366530.8964861741.13512629541.13512629548.84346180678.8434618067

7.927200445828.7954611741.54442810371.54442810373.83564857893.8356485789

6.3458798142.76384933352.7638493335

3%

20%



00

00

00

00

00

00

00

00

00

00

0

Embryo 6- 3% O2

Embryo 6-20% O2

Days


Embryo 5-20% O2DATA


11910000123-May-9800

27720000226-May-9831.2187538443

88703999.9999996329-May-9862.8968257494

217029119.99999942-Jun-98104.1876433117

194602777.59999956-Jun-98144.0302869838

423930214.399997613-Jun-98215.1535814757

2205850215.59465725-Jun-98337.5330176071

21911445474.9069811-Jul-984910.845295532

69532320307.0378916-Jul-985412.5112946048

4422255571527.6108-Aug-987718.5022494652

57430359022238.51115-Aug-988422.2012087368

Embryo 5-3% O2DATA


11910000123-May-9800

27720000226-May-9831.2187538443

120397200329-May-9863.3375565223

551017851.99999742-Jun-98105.531853742

2235295752.9466556-Jun-98147.5521485042

6981573735.036769-Jun-98179.1952350514

29485513074.305713-Jun-982111.27361717

102314730367.838816-Jun-982413.0685528328

411987314281.162920-Jun-982815.0781388817

2709503236922.441026-Jun-983417.7954954319

13755244766109.61130-Jun-983820.1393769679



200000012-Jun-9805.531853742

264000023-Jun-9815.9323916716

555000039-Jun-9877.0043415135

47767000.0000001416-Jun-981410.1097961083

169413626.666667520-Jun-981811.9362598533

1212436854.84444626-Jun-982414.7755476386

5472131671.53123730-Jun-982816.9497409717

Embryo 6-20% O2DATA


12020000123-May-9800

30020000226-May-9831.3204870809

121781133.333333329-May-9863.3407818431

375897764.88888842-Jun-98104.9668315359

626496274.81481356-Jun-98145.7037971301

1585035575.28148625-Jun-98337.042934515

3138746336.82221711-Jul-98498.0286077335

6244079538.98765816-Jul-98549.0209082086

182077359356.8898-Aug-987713.8868270232

1525808271410.651015-Aug-988416.9537772671

Embryo 6- 3% O2DATA


12020000123-May-9800

30020000226-May-9831.3204870809

148799133.333333329-May-9863.6298573225

691419972.88888742-Jun-98105.8460534778

3072209412.8696256-Jun-98147.9976958524

9155184050.3514469-Jun-98179.5730081831

30730901129.013713-Jun-982111.3200374606

88095249903.1704816-Jun-982412.8394116197

287777816350.357920-Jun-982814.5472308682

2497911445921.091026-Jun-983417.6649259109

16877555002940.21130-Jun-983820.4212375939



200000012-Jun-9805.8460534778

253200023-Jun-9816.1863308826

558000039-Jun-9877.3263185999

49178400416-Jun-981410.4660063709

156387312520-Jun-981812.1350331364

1091583437.76626-Jun-982414.9382601728

5905466398.28157730-Jun-982817.3738887669



119100001 (thawed)23-May-9800000119100002 in 3% O2


88703999.9999998329-May-98631.787766061196000002.89682574941.678071905130000003166000005A-20%

217029119.99999942-Jun-981043.098811262573400004.18764331171.290817562330000003143400005B-20%

194602777.59999956-Jun-98144-25.420013623826900004.0302869838-0.1573563279269000030

423930214.399998613-Jun-982176.231669478258600005.15358147571.12329449199000003149600005C-20%(senesc)

2205850215.59466725-Jun-9833125.043211642146830007.53301760712.37943613156000003140830005D-20%

21911445474.9069811-Jul-9849164.8305125243596000010.8452955323.312277924915000003144600005E - 20%

69532320307.0379916-Jul-985453.0012021505476000012.51129460481.66599907281500003146100005F-20%

4422255571527.61108-Aug-9877233.8391208974954000018.50224946525.990954860415000003180400005G-20%

57430359022238.61115-Aug-988471.89242418911948000022.20120873683.69895927164000000Frozen1154800005H-20%



119100001 (thawed)23-May-9800000119100002 in 3% O2


120397200329-May-98631.4158940004130300003.33755652232.1188026781300000031100300005A-3%

55101785242-Jun-981041.8229071086137300005.5318537422.194297219730000003 and UV (1)177300005B-3%

2235295752.9466756-Jun-981441.9799091077121700007.55214850422.020294762230000003191700005C-3%

6981573735.0367569-Jun-981731.825832002193700009.19523505141.643086547230000003163700005D-3%

29485513074.3052713-Jun-982141.92457390971267000011.273617172.078382118630000003196700005E - 3%

102314730367.839816-Jun-982431.67136909821041000013.06855283281.794935662830000003174100005F-3%

411987314281.164920-Jun-982841.99045967821208000015.07813888172.009586048830000003190800005G-3%

2709503236922.451026-Jun-983462.20802823961973000017.79549543192.7173565502300000031167300005H-3%

13755244766109.61130-Jun-983841.70657089051523000020.13937696792.3438815364000000Frozen1112300005I-3%



20000001 (see 3% O2)2-Jun-980005.531853742020000001


555000039-Jun-98765.59727681855500007.00434151351.071949841915000003140500005A-UV

47767000416-Jun-981472.25409832481291000010.10979610833.105454594830000003199100005B-UV

169413626.666667520-Jun-981842.19002430841064000011.93625985331.82646374530000003176400005C-UV

1212436854.84444626-Jun-982462.11320600572147000014.77554763862.8392877854300000031184700005D-UV

5472131671.53125730-Jun-982841.83976279351354000016.94974097172.17419333314000000Frozen195400005E - UV



120200001 (thawed)23-May-9800000120200002 in 3% O2


121781133.333334329-May-98631.4849318308121700003.34078184312.020294762230000003191700006A-20%

375897764.88888942-Jun-981042.459949420992600004.96683153591.626049692830000003162600006B-20%

626496274.81481656-Jun-981445.427661795450000005.70379713010.73696559423000003147000006C-20%(senesc)

1585035575.28148625-Jun-98331914.18823805087590007.0429345151.339137384975900030

3138746336.82223711-Jul-98491616.232560345215030008.02860773350.9856732185150300030

6244079538.98766816-Jul-985455.038796337529900009.02090820860.99230047521500003128400006D-20%

182077359356.8898-Aug-9877234.7267537492437400013.88682702324.865918814615000003128740006E - 20%

1525808271410.661015-Aug-988472.28239764041257000016.95377726713.06695024394000000Frozen185700006F-20%

Embryo 6- 3% O2DATADATADATA


120200001 (thawed)23-May-9800000120200002 in 3% O2


148799133.333334329-May-98631.2990554507148700003.62985732252.3093702416300000031118700006A-3%

691419972.8888942-Jun-981041.8048943864139400005.84605347782.216196155430000003 and UV (1)179400006B-3%

3072209412.8696356-Jun-981441.8590450008133300007.99769585242.1516423746300000031103300006C-3%

9155184050.3515169-Jun-981731.904384255589400009.57300818311.575312330730000003159400006D-3%

30730901129.0132713-Jun-982142.2896010111007000011.32003746061.747029277530000003170700006E - 3%

88095249903.1712816-Jun-982431.9744971849860000012.83941161971.519374159130000003156000006F-3%

287777816350.359920-Jun-982842.3421682379980000014.54723086821.707819248530000003168000006G-3%

2497911445921.121026-Jun-983461.92449868182604000017.66492591093.1176950427300000031230400006H-3%

16877555002940.31130-Jun-983841.45121468842027000020.42123759392.7563116834000000Frozen1162700006I-3%

Embryo 6- 3% + UVDATADATADATA


20000001 (see 3% O2)2-Jun-980005.8460534778020000001


558000039-Jun-98765.263214602255800007.32631859991.139987717315000003140800006A-UV

49178400416-Jun-981472.22952105771322000010.46600637093.139687771300000031102200006B-UV

156387312520-Jun-981842.3966062634954000012.13503313641.669026765530000003165400006C-UV

1091583437.76626-Jun-982462.14039031522094000014.93826017282.8032270364300000031179400006D-UV

5905466398.28159730-Jun-982841.64228651681623000017.37388876692.43562859414000000Frozen1122300006E - UV

&F

Page &P

Chart1

8.85798174493.74003673675.1179450082

11.4468976485.86304513685.5838525112

8.47950825873.01493626985.4645719889

23.458915893910.612366713912.84654918

32.064525246223.86197227638.20255297

3%O2

20%O2

total

no-change

size-change


Mutant Frequency (x10 -5)

Sheet1

Sheet1

8.85798174493.74003673675.1179450082

11.4468976485.86304513685.5838525112

8.47950825873.01493626985.4645719889

23.458915893910.612366713912.84654918

32.064525246223.86197227638.20255297

3%O2

20%O2

total

no-change

size-change


Mutant Frequency (x10 -5)

Sheet2

Sheet3

MBD04DB7550.xls

Chart8

9.2431113868.6599233282.29180274962.29180274961.84988161521.8498816152

9.695682944111.5450952211.52820506961.52820506964.11345175544.1134517554

14.044824660618.50027367270.90205882840.90205882844.24737156894.2473715689

17.969602915425.02284362021.66227392291.66227392295.62109280885.6210928088

10.49976072333.83028598224.34485506164.34485506165.35562804185.3556280418

18.609022556433.45863503950.27873914830.27873914831.44869034531.4486903453

14.57471264372.57798123572.5779812357

11.96507368832.87247411682.8724741168

13.11611313274.02038027144.0203802714

3%O2

20%O2



Sheet3



Embryo 5-3% O226-May-9831.2187538443

Embryo 5-3% O229-May-9863.3375565223

Embryo 5-3% O22-Jun-98105.531853742







Embryo 5-3% O230-Jun-983820.1393769679


Embryo 5-3% + UV26-May-9831.2187538443

Embryo 5-3% + UV29-May-9863.3375565223

Embryo 5-3% + UV2-Jun-98105.531853742

Embryo 5-3% + UV3-Jun-98115.9323916716

Embryo 5-3% + UV9-Jun-98177.0043415135

Embryo 5-3% + UV16-Jun-982410.1097961083

Embryo 5-3% + UV20-Jun-982811.9362598533

Embryo 5-3% + UV26-Jun-983414.7755476386

Embryo 5-3% + UV30-Jun-983816.9497409717

Embryo 6- 3% O223-May-9800

Embryo 6- 3% O226-May-9831.3204870809

Embryo 6- 3% O229-May-9863.6298573225

Embryo 6- 3% O22-Jun-98105.8460534778

Embryo 6- 3% O26-Jun-98147.9976958524

Embryo 6- 3% O29-Jun-98179.5730081831

Embryo 6- 3% O213-Jun-982111.3200374606

Embryo 6- 3% O216-Jun-982412.8394116197

Embryo 6- 3% O220-Jun-982814.5472308682

Embryo 6- 3% O226-Jun-983417.6649259109

Embryo 6- 3% O230-Jun-983820.4212375939

Embryo 6- 3% + UV23-May-9800

Embryo 6- 3% + UV26-May-9831.3204870809

Embryo 6- 3% + UV29-May-9863.6298573225

Embryo 6- 3% + UV2-Jun-98105.8460534778

Embryo 6- 3% + UV3-Jun-98116.1863308826

Embryo 6- 3% + UV9-Jun-98177.3263185999

Embryo 6- 3% + UV16-Jun-982410.4660063709

Embryo 6- 3% + UV20-Jun-982812.1350331364

Embryo 6- 3% + UV26-Jun-983414.9382601728

Embryo 6- 3% + UV30-Jun-983817.3738887669

Sheet3

0000

0000

0000

0000

0000

0000

0000

0000

0000

0000

0000

0000

Embryo 5-3% + UV

Embryo 5-3% O2

Embryo 6- 3% + UV

Embryo 6- 3% O2

days


Sheet2

Cell typeCumm cell#PassageDateDay# totalPD#totalCell typeCumm cell#PassageDateDay# totalPD#totalSum of PD#totalDay# total

Embryo 5-20% O211910000123-May-9800Embryo 5-20% O211910000123-May-9800Cell type03610141721242833343849547784

Embryo 5-20% O227720000226-May-9831.2187538443Embryo 5-20% O227720000226-May-9831.2187538443Embryo 5-20% O201.21875384432.89682574944.18764331174.03028698385.15358147577.533017607110.84529553212.511294604818.502249465222.2012087368

Embryo 5-20% O288703999.9999996329-May-9862.8968257494Embryo 5-20% O288703999.9999996329-May-9862.8968257494Embryo 5-3% O201.21875384433.33755652235.5318537427.55214850429.195235051411.2736171713.068552832815.078138881717.795495431920.1393769679

Embryo 5-20% O2217029119.99999942-Jun-98104.1876433117Embryo 5-20% O2217029119.99999942-Jun-98104.1876433117Embryo 6- 3% O201.32048708093.62985732255.84605347787.99769585249.573008183111.320037460612.839411619714.547230868217.664925910920.4212375939

Embryo 5-20% O2194602777.59999956-Jun-98144.0302869838Embryo 5-20% O2194602777.59999956-Jun-98144.0302869838Embryo 6-20% O201.32048708093.34078184314.96683153595.70379713017.0429345158.02860773359.020908208613.886827023216.9537772671





Embryo 5-20% O24422255571527.6108-Aug-987718.5022494652Embryo 5-20% O24422255571527.6108-Aug-987718.5022494652MEF 5

Embryo 5-20% O257430359022238.51115-Aug-988422.2012087368Embryo 5-20% O257430359022238.51115-Aug-988422.2012087368




















Embryo 6-20% O230020000226-May-9831.3204870809Embryo 6-20% O2182077359356.8898-Aug-987713.8868270232

Embryo 6-20% O2121781133.333333329-May-9863.3407818431Embryo 6-20% O21525808271410.651015-Aug-988416.9537772671

Embryo 6-20% O2375897764.88888842-Jun-98104.9668315359Embryo 6-3% O212020000123-May-9800

Embryo 6-20% O2626496274.81481356-Jun-98145.7037971301Embryo 6-3% O230020000226-May-9831.3204870809

Embryo 6-20% O21585035575.28148625-Jun-98337.042934515Embryo 6-3% O2148799133.333333329-May-9863.6298573225





Embryo 6- 3% O212020000123-May-9800Embryo 6-3% O288095249903.1704816-Jun-982412.8394116197Sum of Cumm cell#Day# total

Embryo 6- 3% O230020000226-May-9831.3204870809Embryo 6-3% O2287777816350.357920-Jun-982814.5472308682Cell type03610141721242833343849547784

Embryo 6- 3% O2148799133.333333329-May-9863.6298573225Embryo 6-3% O22497911445921.091026-Jun-983417.6649259109Embryo 5-20% O2119100002772000088703999.9999996217029119.999999194602777.599999423930214.3999972205850215.5946521911445474.906969532320307.03784422255571527.657430359022238.5

Embryo 6- 3% O2691419972.88888742-Jun-98105.8460534778Embryo 6-3% O216877555002940.21130-Jun-983820.4212375939Embryo 5-3% O21191000027720000120397200551017851.9999972235295752.946656981573735.036729485513074.305102314730367.838411987314281.1622709503236922.4413755244766109.6

Embryo 6- 3% O23072209412.8696256-Jun-98147.9976958524Embryo 6-20% O21202000030020000121781133.333333375897764.888888626496274.8148131585035575.281483138746336.822216244079538.98765182077359356.881525808271410.65

Embryo 6- 3% O29155184050.3514469-Jun-98179.5730081831Embryo 6-3% O21202000030020000148799133.333333691419972.8888873072209412.869629155184050.3514430730901129.01388095249903.1704287777816350.3572497911445921.0916877555002940.2

Embryo 6- 3% O230730901129.013713-Jun-982111.3200374606

Embryo 6- 3% O288095249903.1704816-Jun-982412.8394116197

Embryo 6- 3% O2287777816350.357920-Jun-982814.5472308682

Embryo 6- 3% O22497911445921.091026-Jun-983417.6649259109

Embryo 6- 3% O216877555002940.21130-Jun-983820.4212375939

Embryo 6- 3% + UV200000012-Jun-9805.8460534778

Embryo 6- 3% + UV253200023-Jun-9816.1863308826

Embryo 6- 3% + UV558000039-Jun-9877.3263185999

Embryo 6- 3% + UV49178400416-Jun-981410.4660063709

Embryo 6- 3% + UV156387312520-Jun-981812.1350331364

Embryo 6- 3% + UV1091583437.76626-Jun-982414.9382601728

Embryo 6- 3% + UV5905466398.28157730-Jun-982817.3738887669

MEF 6

bold 3%

open 20%

Sheet2

0000

0000

0000

0000

0000

0000

0000

0000

0000

0000

0000

0000

0000

0000

0000

0000

Embryo 5-20% O2

Embryo 5-3% O2

Embryo 6- 3% O2

Embryo 6-20% O2

Days


Sheet1

0000

0000

0000

0000

0000

0000

0000

0000

0000

0000

0000

0000

0000

0000

0000

0000

Embryo 5-20% O2

Embryo 5-3% O2

Embryo 6-20% O2

Embryo 6-3% O2

Days

Cummulative Cell Number

PDWorksheet

00

00

00

00

00

00

00

00

00

00

00

20%

3%

Days


00

00

00

00

00

00

00

00

00

00

0

Embryo 6- 3% O2

Embryo 6-20% O2

Days


9.2431113868.6599233282.29180274962.29180274961.84988161521.8498816152

9.695682944111.5450952211.52820506961.52820506964.11345175544.1134517554

14.044824660618.50027367270.90205882840.90205882844.24737156894.2473715689

17.969602915425.02284362021.66227392291.66227392295.62109280885.6210928088

10.49976072333.83028598224.34485506164.34485506165.35562804185.3556280418

18.609022556433.45863503950.27873914830.27873914831.44869034531.4486903453

14.57471264372.57798123572.5779812357

11.96507368832.87247411682.8724741168

13.11611313274.02038027144.0203802714

3%O2

20%O2



9.2431113868.6599233282.29180274962.29180274961.84988161521.8498816152

9.695682944111.5450952211.52820506961.52820506964.11345175544.1134517554

14.044824660618.50027367270.90205882840.90205882844.24737156894.2473715689

17.969602915425.02284362021.66227392291.66227392295.62109280885.6210928088

10.49976072333.83028598224.34485506164.34485506165.35562804185.3556280418

18.609022556433.45863503950.27873914830.27873914831.44869034531.4486903453

14.57471264372.57798123572.5779812357

11.96507368832.87247411682.8724741168

13.11611313274.02038027144.0203802714

3%O2

20%O2



10.56940046347.90559824652.06045891382.06045891381.03263062931.0326306293

6.37332768549.5047341791.12823378641.12823378642.29525595642.2952559564

11.58867707413.40901696895.23048614315.23048614314.56858980924.5685898092

10.637889688213.73278716223.28489319463.28489319462.74780256512.7478025651

11.09791466513.35719502010.71386190940.71386190945.89270276815.8927027681

9.31682864918.79208982010.6790505560.6790505561.90940713411.9094071341

9.942993366530.8964861741.13512629541.13512629548.84346180678.8434618067

7.927200445828.7954611741.54442810371.54442810373.83564857893.8356485789

6.3458798142.76384933352.7638493335

3%

20%



10.56940046347.90559824652.06045891382.06045891381.03263062931.0326306293

6.37332768549.5047341791.12823378641.12823378642.29525595642.2952559564

11.58867707413.40901696895.23048614315.23048614314.56858980924.5685898092

10.637889688213.73278716223.28489319463.28489319462.74780256512.7478025651

11.09791466513.35719502010.71386190940.71386190945.89270276815.8927027681

9.31682864918.79208982010.6790505560.6790505561.90940713411.9094071341

9.942993366530.8964861741.13512629541.13512629548.84346180678.8434618067

7.927200445828.7954611741.54442810371.54442810373.83564857893.8356485789

6.3458798142.76384933352.7638493335

3%

20%



00

00

00

00

00

00

00

00

00

00

0

Embryo 6- 3% O2

Embryo 6-20% O2

Days


Embryo 5-20% O2DATA


11910000123-May-9800

27720000226-May-9831.2187538443

88703999.9999996329-May-9862.8968257494

217029119.99999942-Jun-98104.1876433117

194602777.59999956-Jun-98144.0302869838

423930214.399997613-Jun-98215.1535814757

2205850215.59465725-Jun-98337.5330176071

21911445474.9069811-Jul-984910.845295532

69532320307.0378916-Jul-985412.5112946048

4422255571527.6108-Aug-987718.5022494652

57430359022238.51115-Aug-988422.2012087368

Embryo 5-3% O2DATA


11910000123-May-9800

27720000226-May-9831.2187538443

120397200329-May-9863.3375565223

551017851.99999742-Jun-98105.531853742

2235295752.9466556-Jun-98147.5521485042

6981573735.036769-Jun-98179.1952350514

29485513074.305713-Jun-982111.27361717

102314730367.838816-Jun-982413.0685528328

411987314281.162920-Jun-982815.0781388817

2709503236922.441026-Jun-983417.7954954319

13755244766109.61130-Jun-983820.1393769679



200000012-Jun-9805.531853742

264000023-Jun-9815.9323916716

555000039-Jun-9877.0043415135

47767000.0000001416-Jun-981410.1097961083

169413626.666667520-Jun-981811.9362598533

1212436854.84444626-Jun-982414.7755476386

5472131671.53123730-Jun-982816.9497409717

Embryo 6-20% O2DATA


12020000123-May-9800

30020000226-May-9831.3204870809

121781133.333333329-May-9863.3407818431

375897764.88888842-Jun-98104.9668315359

626496274.81481356-Jun-98145.7037971301

1585035575.28148625-Jun-98337.042934515

3138746336.82221711-Jul-98498.0286077335

6244079538.98765816-Jul-98549.0209082086

182077359356.8898-Aug-987713.8868270232

1525808271410.651015-Aug-988416.9537772671

Embryo 6- 3% O2DATA


12020000123-May-9800

30020000226-May-9831.3204870809

148799133.333333329-May-9863.6298573225

691419972.88888742-Jun-98105.8460534778

3072209412.8696256-Jun-98147.9976958524

9155184050.3514469-Jun-98179.5730081831

30730901129.013713-Jun-982111.3200374606

88095249903.1704816-Jun-982412.8394116197

287777816350.357920-Jun-982814.5472308682

2497911445921.091026-Jun-983417.6649259109

16877555002940.21130-Jun-983820.4212375939



200000012-Jun-9805.8460534778

253200023-Jun-9816.1863308826

558000039-Jun-9877.3263185999

49178400416-Jun-981410.4660063709

156387312520-Jun-981812.1350331364

1091583437.76626-Jun-982414.9382601728

5905466398.28157730-Jun-982817.3738887669



119100001 (thawed)23-May-9800000119100002 in 3% O2


88703999.9999998329-May-98631.787766061196000002.89682574941.678071905130000003166000005A-20%

217029119.99999942-Jun-981043.098811262573400004.18764331171.290817562330000003143400005B-20%

194602777.59999956-Jun-98144-25.420013623826900004.0302869838-0.1573563279269000030

423930214.399998613-Jun-982176.231669478258600005.15358147571.12329449199000003149600005C-20%(senesc)

2205850215.59466725-Jun-9833125.043211642146830007.53301760712.37943613156000003140830005D-20%

21911445474.9069811-Jul-9849164.8305125243596000010.8452955323.312277924915000003144600005E - 20%

69532320307.0379916-Jul-985453.0012021505476000012.51129460481.66599907281500003146100005F-20%

4422255571527.61108-Aug-9877233.8391208974954000018.50224946525.990954860415000003180400005G-20%

57430359022238.61115-Aug-988471.89242418911948000022.20120873683.69895927164000000Frozen1154800005H-20%



119100001 (thawed)23-May-9800000119100002 in 3% O2


120397200329-May-98631.4158940004130300003.33755652232.1188026781300000031100300005A-3%

55101785242-Jun-981041.8229071086137300005.5318537422.194297219730000003 and UV (1)177300005B-3%

2235295752.9466756-Jun-981441.9799091077121700007.55214850422.020294762230000003191700005C-3%

6981573735.0367569-Jun-981731.825832002193700009.19523505141.643086547230000003163700005D-3%

29485513074.3052713-Jun-982141.92457390971267000011.273617172.078382118630000003196700005E - 3%

102314730367.839816-Jun-982431.67136909821041000013.06855283281.794935662830000003174100005F-3%

411987314281.164920-Jun-982841.99045967821208000015.07813888172.009586048830000003190800005G-3%

2709503236922.451026-Jun-983462.20802823961973000017.79549543192.7173565502300000031167300005H-3%

13755244766109.61130-Jun-983841.70657089051523000020.13937696792.3438815364000000Frozen1112300005I-3%



20000001 (see 3% O2)2-Jun-980005.531853742020000001


555000039-Jun-98765.59727681855500007.00434151351.071949841915000003140500005A-UV

47767000416-Jun-981472.25409832481291000010.10979610833.105454594830000003199100005B-UV

169413626.666667520-Jun-981842.19002430841064000011.93625985331.82646374530000003176400005C-UV

1212436854.84444626-Jun-982462.11320600572147000014.77554763862.8392877854300000031184700005D-UV

5472131671.53125730-Jun-982841.83976279351354000016.94974097172.17419333314000000Frozen195400005E - UV



120200001 (thawed)23-May-9800000120200002 in 3% O2

30020000226-May-98332.2718889441300200001.32048708091.

genomic instability in cancer and aging

Documents

genome deterioration

percentageoymutant frequencies

micemutant frequency

genomic instability

mouse organsmutant spectra

vitropoint mutational

san antonio

agingjan vijg