optia – efficiency considerations with mnc collection...manzone et.al. • journal of nuclear...
TRANSCRIPT
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Optia – Efficiency Considerations With MNC Collection
Dr. Volker Witt
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OPTIA™ system (CaridianBCT, Lakewood, Dnv, CO, USA)
• Introduced as the replacement for the COBE spectra system
• TPE, RCE available all over the world
• MNZ, Granulo, Leukodepletion are available in Europe
• BM processing is pending • Universal apheresis system
for therapies and collection of blood and tissue products
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Harvesting autologous PBMNC in children
• Routine procedure in many pediatric oncology therapy protocols – Neuroblastoma
– Ewing sarcoma
– Nephroblastoma
– Hepatoblastoma
– Medulloblastoma
– CNS tumours
– Lymphoma
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Autologous PBCS harvesting in children
Some of the problems • Extracorporeal blood volume
of the apheresis systems • Difficult venous access • Different physiological aspects
in children • Apheresis procedures in
children are accounting for less than 5% of the total number of procedures performed
• Only a few centres are specialized in pediatric apheresis.
Systems in use
• Fenwal CS 3000
– expired
• COBE spectra
– will expire 2014 in Europe
• AMICUS
– still in use
• Fresenius COMTTEC
– rarely used
• Optia
– No published data in pediatrics
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Apheresis procedures in the St. Anna Kinderspital from 1996 to QI-2013
0
50
100
150
200
250
300
350
AUTO DC FD DLI ERY ECP RCE TPE IA WBC XX
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The use of the apheresis systems in children at St. Anna Kinderspital in 2011+2012
OPTIASpectra
AMICUSothers
0
20
40
60
80
100
120
OPTIA
Spectra
AMICUS
others
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collection efficiency : What is efficient?
• Which cell type do I want to collect (HPC, MNC, monocytes, CD3+ T-cells,…)?
• Collecting enough cells for the poposed therapy?
• Collecting enough cells per kg bodyweight?
• Collecting all cells which are running into the apheresis system?
• Collecting all cells per liter whole blood running into the apheresis system?
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Factors influencing the calculation of the collection efficiency
• Cell number of the target cell population
• Total cell number
– Cells who can interfere with the target cell population
• Relation between the target cell population and the cells of no interest
• A sufficient and continous blood flow for optimal separation and harvesting
• Cell loss in the set
• A constant and valid method to describe and enumerate the target cell population
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Model of apheresis
BM PB AP
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Recruitment phenomenon
• The number of cells mobilized into the peripheral blood is even higher than the number of collected cells.
[Mayer J. Bone Marrow Transplantation, 2003, 749-757]
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Yield of a cell population
cells in the product [/ml] X harvest volume [ml]
[ cells x 10E06]
[ cells x 10E06/kg bw]
[Strasser E.F. Transfusion 2005, 445-452]
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Cell loss
(1-(postdonation cell count/predonation cell count))X(-100)[%]
pre- or postdonation cell count =
absolute number of cells in blood pre- and postdonation =
blood cell count [x10E09/L] x total blood volume [L]
[Strasser E.F. Transfusion 2005, 445-452]
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Cell yield per liter of processed blood volume (/L PBV)
Y ÷ PBV [/L]
Y = Yield
PBV = processed blood volume [L]
[Strasser E.F. Transfusion 2005, 445-452]
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Recruitment factor (RF)
(postdonation cell count + cell yield) ÷ predonation cell count
pre- or postdonation cell count =
absolute number of cells in blood pre- and postdonation =
blood cell count [x10E09/L] x total blood volume [L]
[Strasser E.F. Transfusion 2005, 445-452]
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Collection Efficiency (CE)
Efficiency of the procedure [%]
= cells in the harvest ÷ cells processed
Witt V. Journal of Clinical Apheresis; 2001; 161-168
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Collection Efficiency (CE)
a = cells/mL
b = harvest volume in mL
c = cells/mL before harvest
d = cells/mL after harvest
e = processed blood volume [mL]
absolute cell count in the harvest = a * b
processed blood volume (e) = whole blood volume processed – ACD-A volume used
Witt V. Journal of Clinical Apheresis; 2001; 161-168
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Manzone et.al. • JOURNAL OF NUCLEAR MEDICINE TECHNOLOGY • 2007 • 55-63
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Blood volume is the volume of blood (both red blood cells and plasma) in a person's circulatory system. A typical adult has a blood volume of approximately between 4.7 and 5 liters, with females generally having less blood volume than males. Blood volume is regulated by the kidneys. Blood volume (BV) can be calculated given the hematocrit (HCT; the fraction of blood that is red blood cells) and plasma volume (PV):
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Total Body Volume (TBV)
Weight in kg x BV in mL/kg = TBV
Patient Fat Thin Normal Muscular
Male 60 65 70 75
Female 55 60 65 70
Infant/Child - - 80/70 -
G. Herold Innere Medizin 2007
Tronier W., Goodfellow E., Larson K. Apheresis Math and Useful Physical constants in Principles of Apheresis Technology, 4th edition Linz W. (editor) 2010
Normal BV = 70 mL/kg bw or 1/14 part of the bodyweight
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Collection efficiency
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Collection efficiency
• Harvested cells (HPC harvested) are calculated by the formula:
Cellsharvested = cell countbuffy [cells/ml] x volumebuffy [ml]
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Collection efficiency
• The cells processed (HPC processed) are calculated by the forumla:
cellsstart – cellsend
2
• The processed blood volume is calculated by the formula:
Inlet volumeproc.– ACD-A used = blood volumeproc.
X processed blood volume
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Collection efficiency st
art
Aft
er p
rim
ing
en
d
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Collection efficiency
• The cells processed are calculated by the formula:
cellsstart – cellsend
2
• This formula describes the mean of cells present during the procedure.
• The cell yield is represented by the area under the curve.
• This approach reflects only an approximation to the real conditions during an apheresis procedure.
X processed blood volume
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Collection efficiency st
art
Aft
er p
rim
ing
en
d
A
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Collection efficiency st
art
Aft
er p
rim
ing
en
d
B
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Collection efficiency st
art
Aft
er p
rim
ing
en
d
C
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Collection efficiency
•A > B
•A < C
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Collection ratio
• CR = ((CD34+ /µl product x PV) /kg donor BW) / CD34+ pre
• This formular gives an idea about the collection efficiency of one cell populationen independently from the donor or patients weight.
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Collection efficiency
• Ideal times points for target cell count to calculate the „true“ collection efficiency:
Immediately before start of the apheresis
End of priming (saline and blood)
Before reinfusion
Immediately after end of the apheresis
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Collection efficiency
• For practical reasons target cell counts are performed only before and after apheresis procedure
• For economic reasons target cell count are performed before apheresis procedure only
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Collection efficiency
• How good could the target cell yield predicted by measuring the target cell population before the apheresis procedure only?
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N = 7 Median Min - Max
Bodyweight 35 kg 10 – 55
Bodyweight < 20 kg 3/7 10.25, 12.7 and 19 kg
Body lenght 1.45 m 0.75 – 1.72
Body surface 1.19 m² 0.46 – 1.61
Age 11.8 years 1.03 - 20
Sex 2 female 5 male
Underlyingdiseases neuroblastoma, hepatoblastoma, ATRT, PNET, AML and 2 Ewing’s sarcoma
Patients_AUTO
N = 16 Median Min - Max
Bodyweight 30,50 8 – 89
Bodyweight < 20 kg 7/16 12.70, 10.25, 19, 17, 10.7, 12.7 and 8 kg
Body lenght (cm) 134 72 – 187
Body surface (m²) 1,07 0.4 – 2.17
Age 9,88 1,03 - 20,03
Sex 4 female 12 male
Underlying diseases 2 ATRT, AML recurrency, Hepatoblastoma, 3 Neuroblastoma, PNET, 5 Ewing's sarcoma, Retinoblastoma recurrency, germ cell tumor and Rhabdoid tumor
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N = 9 Median Min - Max
Bodyweight (kg) 36 27.4 – 45
Bodyweight < 20 kg 0/9 -
Body lenght (cm) 130 125 - 158
Body surface (m²) 1,12 0.99 - 1.41
Age (years) 8,65 7.12 - 20.13
Sex 3 female 6 male
Underlying diseases 8 GVHD and ALL/GVHD
Patients_ECP
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Parameter settings_AUTO
N=16 Median Min – Max
Total whole blood drawn (ml) 5267 1687 - 18882
Whole blood (ml/kg bw) 211 123 - 322
Time used [hh:mm] 4:15 1.44 - 6:34
Product volume (ml) 90 31 - 332
N cycles 4 2 - 20
ACD-A ratio 13 12 - 15
Collection preferency 30 20 - 40
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Parameter settings_ECP
N=9 Median Min – Max
Total whole blood drawn 3617 2328 - 5100
Whole blood / kg bw 98 58 - 158
Time used 3:04 2:00 - 4:23
Product volume 54 18 - 72
N cycles 3 1 - 4
ACD-A ratio 12 12
Collection preferency 30 20 - 40
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N=16 Median Min - Max
Hemoglobin (g/dl) 10,5 8 - 12
Hematocrit (%) 30,80 21 - 34
Platelets (G/l) 173 41 - 255
Leukocyte (G/l) 15,5 6 - 74
Lymphocytes (%) 4,35 1 - 15.4
Monocytes (%) 11,75 5.5 - 41
MNC (%) 18,55 7 - 59.8
Granulocytes (%) 81,45 40.2 - 93
CD34+ cells (%) 0,33035 0.05 - 1.39
cells/µl 46,5 18 - 270
Pre apheresis values_AUTO
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Pre apheresis values_ECP
N=9 Median Min - Max
Hemoglobin (g/dl) 11,4 10 - 12.1
Hematocrit (%) 33,00 30 - 36
Platelets (G/l) 158 31 - 362
Leukocyte (G/l) 4,72 1.3 - 7.07
Lymphocytes (%) 31,1 10 - 52.2
Monocytes (%) 6,2 3.7 - 10.3
MNC (%) 38,3 15.7 - 67.8
Granulocytes (%) 61,6 32.5 - 84.5
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N=16 Median Min - Max
Hemoglobin (g/dl) 9,5 8.2 - 12.8
Hematocrit (%) 27 23 - 35.2
Platelets (G/l) 105 30 - 167
Leukocyte (G/l) 18,8 4.9 - 69
Lymphocytes (%) 4,15 1 - 16.8
Monocytes (%) 10 2.5 - 39.6
MNC (%) 14 4 - 44.4
Granulocytes (%) 86 51 - 95.5
CD34+ cells (%) 0,21 0.01 - 1.34
cells/µl 32 0.2 - 194
Post apheresis values_AUTO
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Post apheresis values_ECP
N=9 Median Min - Max
Hemoglobin (g/dl) 10,2 8.6 - 11.6
Hematocrit (%) 29,00 25 - 34
Platelets (G/l) 114 9 - 320
Leukocyte (G/l) 4,85 1.5 - 7.6
Lymphocytes (%) 36,1 8.7 - 59.4
Monocytes (%) 5,3 1.6 - 11.3
MNC (%) 45,7 14.7 - 74.1
Granulocytes (%) 54,4 25.8 - 85.3
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N=16 Median Min - Max
RBC (T/l) 0,25 0.1 - 0.5
Hematocrit (%) 2,65 1 - 7
Platelets (G/l) 1044,5 171 - 2330
Leukocyte (G/l) 154,5 106.7 - 275
Lymphocytes (%) 25,00 4.5 - 48.5
Monocytes (%) 35 5.5 - 58
MNC (%) 68,50 21 - 96.5
Granulocytes (%) 31,50 3.5 - 79
CD34+ cells (%) 0,86 0.35 - 4.18
cells/kg bw 6,1 2.2 - 17
Apheresis product values_AUTO
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Apheresis product values_ECP
N=9 Median Min - Max
RBC (T/l) 0,2 0.1 - 0.5
Hematocrit (%) 2 1 - 6
Platelets (G/l) 563 70 - 2310
Leukocyte (G/l) 50,9 0.9 - 73
Lymphocytes (%) 71,50 6 - 81.5
Monocytes (%) 14 7 - 42.5
MNC (%) 97,50 13.5 - 99.5
Granulocytes (%) 2,50 0.5 - 86.5
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patient Eff_CD34 Eff_lympho Eff_Ery Eff_G Eff_M Eff_Th Eff_MNC Eff_L
AUTO20110001 68,48 57,56 0,1 0,87 8,02 18,9 23,56 7,49
AUTO20110006 67,95 69,64 0,2 7,25 49,46 22,41 59,56 34,75
AUTO20110007 59,56 58,97 0,13 0,63 51,26 23 58,46 13,96
AUTO20110008 26,79 56,34 0,32 5,16 57,86 16,79 55,66 14,76
AUTO20110010 72,38 20,5 0,09 13,75 71,26 18,41 73,35 31,01
AUTO20110011 66,23 66,29 0,19 1,34 55,23 14,65 57,92 6,4
AUTO20110013 83,86 80,31 0,24 11,97 75,64 38,42 79,04 16,06
AUTO20120005 277,38 86,73 0,12 5,13 52,58 11,15 67,41 19,04
AUTO20120008 55,12 50,24 0,12 50,14 54,03 9,56 52,41 51,21
AUTO20120009 74,46 76,54 0,04 6 61,43 1,97 66,63 18,63
AUTO20120008 37,39 60,51 0,1 6,46 25,6 8,23 39 10,14
AUTO20120012 166,94 177,49 0,19 7,91 33,11 17,04 84,15 16,52
AUTO20130001 53,32 132,56 0,24 2,74 22,96 19,79 49,37 10,05
AUTO20130002 117,68 112,41 0,23 3,04 60,86 24,58 87,26 8,61
AUTO20130003 113,92 116,68 0,16 16,03 26,99 19,68 112,16 25,92
AUTO20130004 65,24 59,23 0,07 27,3 2,78 19,6 45,56 29,81
median 68,215 67,965 0,145 6,23 51,92 18,655 59,01 16,29
Collection efficiencies_AUTO
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Collection efficiencies_ECP
patient Eff_lympho Eff_Ery Eff_G Eff_M Eff_Th Eff_MNC Eff_L
ECP20110009 55,81 0,09 8,93 39,55 16,81 52,45 31,46
ECP20110023 32,38 0,14 0,09 16,99 0 26,97 6,62
ECP20110026 21,63 0,05 36,23 64,23 7,92 30,28 35,29
ECP20110031 15,39 0,03 0,15 28,33 0 18,84 4,69
ECP20110056 46,4 0,13 1,15 68,96 18,57 48,45 24,39
ECP20110060 16,62 0,03 0,19 29,39 1,83 19,24 5,41
ECP20110085 66,65 0,19 0,8 56,94 22,46 64,47 46,19
ECP20110088 56,84 0,12 0,2 56,15 29,16 56,48 23,56
ECP20120005 1,93 0,15 0,31 1,5 8,72 0,79 0,61
median 32,38 0,12 0,31 39,55 8,72 30,28 23,56
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HPC IN DETAIL
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Correlation between CD34 positive cell yield and CD34 positive cell number in the peripheral blood_AUTO
y = 15,09x - 20,97 R² = 0,609
0
50
100
150
200
250
300
0 5 10 15 20
CD
34
_P
B_
vo
r
CD 34_10E06_kg
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Correlation between Eff_CD34 and Leukocyte count pre apheresis in peripheral blood_AUTO
y = 0.0019x + 29.404 R² = 0.3182
0
50
100
150
200
250
300
0 20000 40000 60000 80000
Eff
_C
D3
4
Leuko_PB_vor
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Correlation between CE CD34 positive cells and whole blood volume processed_AUTO
y = 0.3507x + 11.216 R² = 0.1306
0
50
100
150
200
250
300
0 100 200 300 400
Eff
_C
D34
WBV_kg BW
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Correlation between CE CD34 positive cells and number of cycles performed_AUTO
y = 2.9646x + 65.498 R² = 0.0814
0
50
100
150
200
250
300
0 5 10 15 20 25
Eff
_C
D3
4
No cycle
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Correlation between Eff_CD34 and collection preference in the peripheral blood_AUTO
0
20
40
60
80
100
120
140
160
180
200
20 30 40
Eff
_C
D3
4
Collection preferency
± Standarddeviation
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IN MONONUCLEAR CELLS =>
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Correlation between pre apheresis MNC and MNC_kg_buffy_AUTO
y = 5,3006x + 2338,6 R2 = 0,215
0
1000
2000
3000
4000
5000
6000
7000
8000
9000
0.00 200.00 400.00 600.00 800.00 1000.00
MN
C_
vo
r
MNC_kg_buffy
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Whole blood processed / kg bw 1 = HPC and 2 = ECP
0
50
100
150
200
250
300
1 2
WB
V_
kg
BW
target
± Standardabweichung
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Number of cycles processed 1 = HPC and 2 = ECP
0
1
2
3
4
5
6
7
0 bis 22 bis 44 bis 8 8 bis12
12 bis15
15 bis20
20 bis25
Nu
mb
er
of
cyc
les
No cycle
1 2
target
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MNC count in blood pre 1 = HPC and 2 = ECP
0
1000
2000
3000
4000
5000
6000
7000
1 2
MN
C_
vo
r
target
± Standardabweichung
U – Test: p = 0.012
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MNC % in the product 1 = HPC and 2 = ECP
0
20
40
60
80
100
120
1 2
MN
C%
_b
uff
y
target
± Standardabweichung
U – Test: p = 0.0035
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MNC content in the product 1 = HPC and 2 = ECP
0
20000
40000
60000
80000
100000
120000
140000
160000
1 2
MN
C_
bu
ffy
target
± Standardabweichung
U – Test: p = 0.001
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MNC content in the product 1 = HPC and 2 = ECP
0
100
200
300
400
500
600
1 2
MN
C_
kg
_b
uff
y
target
± Standardabweichung
U – Test: p = 0.00049
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MNC collection ratio 1 = HPC and 2 = ECP
0
50
100
150
200
250
300
350
400
450
500
1 2
CR
MN
C
target
± Standardabweichung
U – Test: p = 0.95
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CAN WE PREDICT ANYTHING?
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Formel A CD34+ 106/kg = CD34+/µl PB / 10
• Simple formular
• Is only valid in aphereses procedures standardized for blood volume processed, stable collection efficiency (=> parameter settings!)
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Formel A CD34+ 106/kg = CD34+/µl PB / 10
y = 1.509x - 2.0975 R² = 0.6096
0
5
10
15
20
25
30
0 5 10 15 20
Fo
rme
l A
CD 34_10E06_kg
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Formel A CD34+ 106/kg = CD34+/µl PB / 10
y = 7.014ln(x) - 31.585 R² = 0.6052
-15
-10
-5
0
5
10
0 100 200 300
CD
34
/kg
- A
Eff_CD34
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Formel B C = [(AxD):Bx0.5]:1000
• A = CD34+ cells in peripheral blood [1003/ml]
• B = bodyweight [kg]
• C = CD34+ cells/kg bw in the product
• D = processed volume
• Estimation of an efficiency of 50%
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Formel B C = [(AxD):Bx0.5]:1000
y = 1.0115x + 0.1793 R² = 0.5085
0
2
4
6
8
10
12
14
16
18
20
0 5 10 15 20
Fo
rme
l B
CD 34_10E06_kg
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Formel B C = [(AxD):Bx0.5]:1000
y = 5.6977ln(x) - 24.851 R² = 0.6935
-12
-10
-8
-6
-4
-2
0
2
4
6
8
10
0 100 200 300
CD
34
/kg
- B
Eff_CD34
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WE CAN CALCULATE THE BLOOD VOLUME NEEDED TO BE PROCESSED
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D = [(C:0.5)xB:A]x1000
• A = CD34+ cells in peripheral blood [1003/ml]
• B = bodyweight [kg]
• C = CD34+ cells/kg bw in the product
• D = processed volume
• Estimation of an efficiency of 50%
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COLLECTION EFFICIENCY IN PLATELETS =>
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Platelet count in blood pre / CE 1 = HPC and 2 = ECP
0
10
20
30
40
50
60
70
0 100 200 300 400
CE
pla
tele
ts
Platelet count pre
1 2
target
y = -0.0594x + 27.137 R² = 0.1603
0
10
20
30
40
50
60
70
0 200 400
CE
pla
tele
t
Platelet count pre
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Correlation between platelets collection efficiency and number of cycles_AUTO
y = 0.8442x + 11.377 R² = 0.3657
0.00
5.00
10.00
15.00
20.00
25.00
30.00
35.00
40.00
45.00
0 5 10 15 20 25
Eff_
Th
No cycle
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Correlation between platelets collection efficiency and number of cycles_ECP
y = 4.9335x - 2.5341 R² = 0.1888
0.00
5.00
10.00
15.00
20.00
25.00
30.00
35.00
0 1 2 3 4 5
Eff_
Th
No cycle
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Platelet collection ratio versus number of cycle performed
y = 11.336ln(x) + 4.8427 R² = 0.3762
0
5
10
15
20
25
30
35
40
45
50
0 5 10 15 20 25
CR
pla
tele
ts
No cycle
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Side effects
• Hypocalcemia in a 10 kg child (Ca++ 0.86 mmol/l) complete restitutio after Ca 10% i.v.
• Leackage in the set => change of set, procedure was finished with 30 min delay
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Engraftment data
patient disease bw transplanted engrafted
AUTO20110001 ATRT 40,00 y y
AUTO20110006 AML recurrency 54,50 y y
AUTO20110007
Hepatoblastom
/hepatozelluläres
Karzinom 12,70
n n.a.
AUTO20110008 NB 10,25 n n.a.
AUTO20110010 PNET zentral 19,00 n n.a.
AUTO20110011 Ewing Sarkom 35,00 n n.a.
AUTO20110013 Ewing Sarkom 51,30 n n.a.
So far 2 / 7 patients are treated with Mega therapy and autologous rescue, both engrafted regulary.
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Triggering first chamber flush
• „Less operator input was required, although 26% of Spectra Optia apheresis procedures required triggering of the first chamber flush.” [1]
1 Brauninger Transfusion 2011
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• Flow pauses were 40% less frequent with the Spectra Optia (Table 2), likely by virtue of electronic interface management and an algorithm that automatically restarts the flow if pressure recovers quickly.
1 Brauninger Transfusion 2011
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1 Brauninger Transfusion 2011
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1 Brauninger Transfusion 2011
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Allogeneic stem cell apheresis outcomes with the COBE Spectra MNC
and the Spectra Optia v.5.0. (A) Collection efficiency for CD34+ cells is
negatively correlated with preapheresis CD34+ cell counts.
Preapheresis circulating CD34+ cells counts (X-axis) are plotted over
the corresponding collection efficiency CE1 (Y-axis) for this donor. (
,—) Results from the Spectra Optia; (, - - -) data fromthe COBE Spectra
MNC.
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Spectra Optia apheresis procedures are associated with better
collection efficiency in less well-mobilized donors. Preapheresis
circulating CD34+ cell counts were stratified as indicated. CE1
(mean + SEM) for each group is drawn to the Y-axis. Significant
differences between the groups. ( grey) Spectra Optia apheresis
procedures; (black ) COBE Spectra MNC collections.
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• PLT attrition was calculated as (1—postapheresis PLT count/preapheresis PLT count) x 100.
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Spectra Optia PLT attrition correlates with preapheresis PLT counts.
Preapheresis PLT counts (X-axis) are plotted over the corresponding
value for PLT attrition (Y-axis). ( ,—) Results from the Spectra Optia; (, -
- -) data fromthe COBE Spectra MNC.Markedly lower PLT attrition with
the Spectra Optia was observed particularly in donors with fewer PLTs.
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Conclusion
• Safe and efficient for HPC cell harvest in young children and adolescents
• Good correlation of PB HPC cell count and yield and easily predictable
• The same is found for MNC
• The number of cycles processed might be an independent factor for collection efficiency, not only for white blood cells
• Loss of platelets seems to be dependent on the number of cycles necessary