stem cell research solutions and support · 2020-06-16 · 4 life technologies—stem cell...
TRANSCRIPT
Stem cell research solutions and supportMore resources than any other supplier
r&d
live
chat
online forums
field application specialists
web
inar
s
posters
protocols
tech tips
2 life technologies—stem cell solutions
Stem cell research can be time-consuming. Finding support shouldn’t be.
Go to lifetechnologies.com/pscresources to check out protocols, FAQs, publications, scientific posters, videos, and upcoming events.
That’s why Life Technologies gives you easy access to
70+ proven protocols, 2,500+ peer-reviewed publications,
how-to videos, and webinars from leaders in the stem
cell community. And, when you have a question, technical
support is always available by phone, email, live chat, or
in person. It’s all about getting you everything you need to
ensure stem cell success.
Technical resourcesWatch the experts Ask the expertsHands-on training
lifetechnologies.com/stemcell 3
Contents
4 Pluripotent stem cells
4 Culture
8 Tools for reprogramming
10 Detection
11 Analysis
12 Mesenchymal stem cells
12 Culture
14 Differentiation
15 Detection
16 Neural stem cells
16 Culture
16 Differentiation
17 Hematopoietic stem cells
17 Culture
lentivirus—sendai virus—episomal vectors
Field application specialists
4 life technologies—stem cell solutions
Pluripotent stem cells (PSCs)Culture
Pluripotent stem cell (PSC) research requires careful attention to culture conditions. To find the right solution for the right environment, tap into peer-reviewed publications, protocols, instructional videos, and stem cell experts. From KnockOut™ Serum Replacement to Essential 8™ Medium and cell therapy research media, Gibco® products deliver culture with confidence.
lifetechnologies.com/culturepsc KnockOut™ Serum Replacement (KSR)Well-referenced, proven standardKnockout™ serum replacement and basal media deliver confidence, with over 2,400 peer-reviewed publications.
Cat. No. 10828010, 10828028
Data image for KnockOut™ SR XenoFree CTS™ Kit provided by Dr. Hidenori Akutsu, National Center for Child Health and Development, Tokyo, Japan.
Which pluripotent stem cell culture medium is right for you?The standard for serum-free, feeder-based culture of ESCs and iPSCs from multiple species
Feeder-free culture with flexibility to optimize bFGF concentration and maintain differentiation capacity of PSCs
Optimal feeder-free culture of all human ESC and iPSC lines with minimal lot-to-lot variability
Fully documented xeno-free PSC culture reducing the burden in qualifying reagents, facilitating transition, from the bench to the clinic
Product name KnockOut™ Serum Replacement StemPro® hESC SFM Essential 8™ Medium KnockOut™ CTS™ XenoFree ESC/iPSC Kit
cat. no. 10828010, 10828028 a1000701 a14666sa a1448801, a1448701
Recommended applications Multiple species; feeder culture; reprogramming on feeders; differentiation of PSCs
Human; feeder-free culture Human; feeder-free, xeno-free culture; feeder-free reprogramming Human; feeder-based and feeder-free xeno-free culture for cell therapy research
lot-to-lot variability medium medium low medium
cell morphology compact colonies with smooth edges; high nuclear to cytoplasmic ratio
compact colonies with smooth edges; some colonies will have undefined borders with spikey edges
compact colonies with smooth edges; high nuclear to cytoplasmic ratio
compact colonies with smooth edges; some colonies will have undefined borders with spikey edges
components/unit 1 x 100 ml or 500 ml 1 x 100 ml stempro® hesc supplement1 x 500 ml dmem/f12 with Glutamax™ medium1 x 50 ml 25% bsa
1 x 10 ml essential 8™ supplement1 x 500 ml dmem/f-12 (ham) 1:1
1 x 100 ml Knockout™ sr xenofree cts™
1 x 500 ml Knockout™ dmem cts™
1 x 5 ml Knockout™ sr Gf cocktail cts™
additional components required for complete medium
dmem/f12 (1x) with Glutamax™ medium, neaa, bfGf; 2-mercaptoethanol; for a cost-effective option, choose the Knockout™ esc/ipsc media Kit
bfGf and 2-mercaptoethanol no additional components required bfGf cts™ and 2-mercaptoethanol
price/500 ml of complete medium $$ individually $ for kit
$$ $ $$$$
stability of complete medium stable at 4°c for 2 weeks stable at 4°c for 2 weeks stable at 4°c for 2 weeks stable at 4°c for 2 weeks
specifically designed for cell therapy research
no no no Yes
available support resources Protocols for culturing PSCs, publications, hands-on training, product finder
Protocols for culturing PSCs Protocols for culturing PSCs, FAQs, hands-on training Protocols for culturing PSCs, FAQs, product documentation
$—inexpensive $$—moderate $$$—expensive $$$$—exclusive
lifetechnologies.com/stemcell 5
StemPro® hESC SFMFlexible feeder-free culturestempro® hesc sfm optimizes bfGf concentration and maintains the differentiation capability of pscs.
Cat. No. A1000701
Essential 8™ MediumFeeder-free culture for allessential 8™ medium contains only the necessary components for optimal psc culture for every step of your research.
Cat. No. A14666SA
KnockOut™ SR XenoFree CTS™ KitTraceability for cell therapy researchthe Knockout™ sr xenofree cts™ Kit reduces the burden in qualifying reagents, facilitating transition from the bench to the clinic.
Cat. No. A1448801, A1448701Which pluripotent stem cell culture medium is right for you?The standard for serum-free, feeder-based culture of ESCs and iPSCs from multiple species
Feeder-free culture with flexibility to optimize bFGF concentration and maintain differentiation capacity of PSCs
Optimal feeder-free culture of all human ESC and iPSC lines with minimal lot-to-lot variability
Fully documented xeno-free PSC culture reducing the burden in qualifying reagents, facilitating transition, from the bench to the clinic
Product name KnockOut™ Serum Replacement StemPro® hESC SFM Essential 8™ Medium KnockOut™ CTS™ XenoFree ESC/iPSC Kit
cat. no. 10828010, 10828028 a1000701 a14666sa a1448801, a1448701
Recommended applications Multiple species; feeder culture; reprogramming on feeders; differentiation of PSCs
Human; feeder-free culture Human; feeder-free, xeno-free culture; feeder-free reprogramming Human; feeder-based and feeder-free xeno-free culture for cell therapy research
lot-to-lot variability medium medium low medium
cell morphology compact colonies with smooth edges; high nuclear to cytoplasmic ratio
compact colonies with smooth edges; some colonies will have undefined borders with spikey edges
compact colonies with smooth edges; high nuclear to cytoplasmic ratio
compact colonies with smooth edges; some colonies will have undefined borders with spikey edges
components/unit 1 x 100 ml or 500 ml 1 x 100 ml stempro® hesc supplement1 x 500 ml dmem/f12 with Glutamax™ medium1 x 50 ml 25% bsa
1 x 10 ml essential 8™ supplement1 x 500 ml dmem/f-12 (ham) 1:1
1 x 100 ml Knockout™ sr xenofree cts™
1 x 500 ml Knockout™ dmem cts™
1 x 5 ml Knockout™ sr Gf cocktail cts™
additional components required for complete medium
dmem/f12 (1x) with Glutamax™ medium, neaa, bfGf; 2-mercaptoethanol; for a cost-effective option, choose the Knockout™ esc/ipsc media Kit
bfGf and 2-mercaptoethanol no additional components required bfGf cts™ and 2-mercaptoethanol
price/500 ml of complete medium $$ individually $ for kit
$$ $ $$$$
stability of complete medium stable at 4°c for 2 weeks stable at 4°c for 2 weeks stable at 4°c for 2 weeks stable at 4°c for 2 weeks
specifically designed for cell therapy research
no no no Yes
available support resources Protocols for culturing PSCs, publications, hands-on training, product finder
Protocols for culturing PSCs Protocols for culturing PSCs, FAQs, hands-on training Protocols for culturing PSCs, FAQs, product documentation
$—inexpensive $$—moderate $$$—expensive $$$$—exclusive
6 life technologies—stem cell solutions
KnockOut™ ESC/iPSC Media Kit a cost-effective and convenient way to purchase the reagents required for feeder-based culture of human and nonhuman embryonic stem cells (escs) or induced pluripotent stem cells (ipscs). this kit includes dmem/f12 with Glutamax™ medium, Knockout™ serum replacement, and fGf-basic.
Cat. No. A1412901, A1413001
Pluripotent stem cells (PSCs)Complementary culture products
Which extracellular matrix is right for you?we provide extracellular matrices (ecms) that are designed to minimize adaptation time, maximize cell performance, and help meet regulatory requirements. whether you are culturing your cells on feeder layers or require feeder-free, xeno-free conditions for more clinical applications, Gibco® products provide you with all the choices to culture your pscs in the right environment.
Most commonly used basement membrane matrix for ESC and iPSC cultures
Optimized for use with Essential 8™ Medium
Optimal for clinical research applications that require human origin materials only
Product name Geltrex® LDEV-Free hESC Qualified Vitronectin (VTN-N) CELLstart™ CTS™
cat. no. a1413301 a14701sa a1014201
Recommended applications Feeder and feeder-free culture Feeder-free and xeno-free culture Feeder-free, xeno-free culture
recommended medium pairing stempro® hesc sfm essential 8™ medium Knockout™ sr xenofree cts™-supplemented medium
source origin murine human (recombinant) blend of human native proteins
defined no Yes Yes
storage temperature frozen (–20 to –80°c) frozen (–20 to –80° c) refrigerate (2–8°c)
plate preparation time 1 hr incubation at 37°c 1 hr incubation at room temperature 1–2 hrs incubation at 37°c
shelf life of prepared plate 2 weeks at 2–8°c 1 week at 2–8°c 1–7 days at 2–8°c (tightly sealed)
unit size 1 ml or 5 ml 1 ml 2 ml
number of plates coated/unit 16 x 6-well plates from 1 ml vial diluted 1:100
16 x 6-well plates from 1 ml 22 x 6-well plates when diluted 1:50
relative price/6-well plate $ $$ $$$attachment factor (0.1% Gelatin, cat. no. s006100) may be used for feeder-layer attachment.$—inexpensive $$—moderate $$$—expensive $$$$—exclusive
lifetechnologies.com/stemcell 7
Which passaging method is right for you?whether you prefer to passage your cells enzymatically or manually, Gibco® media provides you with choices for psc subculture.
Recommended for clump passaging in feeder and feeder-free cultures
Standard reagent for clump pas-saging in feeder and feeder-free cultures
Quick and gentle dissociation, recommended for single-cell passaging
Recommended for clump passaging with feeder-free Essential 8™ Medium
Optimal for clinical research applications that require human origin materials only
Quick and easy manual passaging tool for feeder and feeder-free cultures
Product name Dispase Collagenase IV Accutase® UltraPure™ 0.5M EDTA
TrypLE™ Select CTS™ StemPro® EZPassage™ Disposable Stem Cell Passaging Tool
cat. no. 17105041 17104019 a1110501 15575020 a1285901 23181010
Recommended applications
Passaging in clumps
Passaging in clumps
Dissociation to single cells
Passaging in clumps
Passaging in clumps or single-cell passaging
Mechanical passaging in clumps
source origin Bacillus polymyxa Clostridium histolyticum
marine chemical (animal origin–free (aof))
recombinant enzyme (aof)
na
form lyophilized powder lyophilized powder liquid, ready to use liquid, dilute 1,000x liquid, ready to use individually wrapped
storage temperature 2–8°c 2–8°c 2–8°c room temperature room temperature room temperature
shelf life of liquid format
2 weeks 2 weeks 2 years 2 years 2 years na
inactivation method inhibition by dilution (no inhibitors needed)
inhibition by dilution (no inhibitors needed)
inhibition by dilution (no inhibitors needed)
inhibition by dilution (no inhibitors needed)
inhibition by dilution (no inhibitors needed)
na
unit size 5 g 1 g 100 ml 4 x 100 ml 100 ml box of 10 units
optimized concentration for use
2 mg/ml 1 mg/ml for feeder-dependent, 10 mg/ml for feeder-free
1x solution 0.5 mm 1x solution na
number of plates passaged/unit
1,250 x 60 mm plates
500 x 60 mm plates at 1 mg/ml, 50 x 60 mm plates at 10 mg/ml
50 x 60 mm plates 50,000 x 60 mm plates (100 ml at 1,000x dilution)
50 x 60 mm plates 10 plates
relative price/60 mm plate
$ $ $$ $ $$ $$$
$—inexpensive $$—moderate $$$—expensive $$$$—exclusive
An iPSC colony cut into evenly sized pieces using the StemPro® EZPassage™ Disposable Stem Cell Passaging Tool.
8 life technologies—stem cell solutions
Tools for reprogrammingSelecting the right reprogramming tool to generate induced pluripotent stem cells (iPSCs) can leave you with questions. And no one has more answers than Life Technologies. From traditional methods (lentivirus) to nonintegrating technologies (Sendai virus and episomal vectors), Invitrogen™ products provide the breadth and depth you depend on. That means more solutions to meet your safety, efficiency, and affordability needs.
lifetechnologies.com/cellreprogram
iPSC colonies generated using Episomal iPSC Reprogramming Vectors Express TRA-1-81 (green) at Day 24.
(A-B) 200,000 primary human fibroblasts were reprogrammed with (A) the CytoTune®-iPS Sendai Reprogramming Kit and (B) lentivirus. Twenty-eight days post-reprogramming, ~245 colonies were observed from CytoTune® reprogramming, while the lentivirus reprogramming generated ~10 colonies. (C-D) 50,000 primary human fibroblasts were reprogrammed with (C) the CytoTune®-iPS Sendai Reprogramming Kit and (D) mRNA. Twenty-eight days post-reprogramming, ~130 iPSC colonies were observed from CytoTune® reprogramming, while 11 iPSC colonies were observed from mRNA reprogramming.
Pluripotent stem cells (PSCs)
A. CytoTune®-iPS Sendai Reprograming Kit B. Lentivirus
C. CytoTune®-iPS Sendai Reprogramming Kit D. mRNA
Lentiviral iPSC reprogramming particlesWell-referenced, flexible technologychoose from eight different high-titer lentiviral ipsc reprogramming particles for your custom ipsc experiments.
lifetechnologies.com/lenti
Episomal iPSC Reprogramming Vectors
Safety with confidencecreate reproducible ipscs from normal and diseased cell types with invitrogen™ episomal vectors.
CytoTune®-iPS Sendai Reprogramming KitHigh efficiency for difficult cell typesreprogram challenging cell types with the easy-to-use, nonintegrating cytotune®-ips sendai reprogramming Kit.
lifetechnologies.com/stemcell 9
Which reprogramming technology is right for you?Well-referenced technology for iPSC generation from fibroblasts
Proven technology for viral-free iPSC generation from normal and diseased cell types
Efficient generation of integration-free iPSCs from multiple somatic cell types
Product name Lentiviral iPSC reprogramming particles
Episomal iPSC Reprogramming Vectors
CytoTune®-iPS Sendai Reprogramming Kit
cat. no. a1357101, a1357201, a1357301, a1357401, a1357501, a1357601, a1357701, a1357801
a14703 a1378001, a1378002
reprogramming efficiency 0.001–0.01% 0.002–0.08% 0.01–1%
Genomic integration-free no Yes Yes
virus-free reprogramming no Yes no
hands-on time one application, selection and Qc of putative colonies required
one application, minimal Qc of putative colonies required
one application, minimal Qc of putative colonies required
blood cell reprogramming no no Yes
convenience hoct4-Gfp and cmv-Gfp reporters available for transduction and expression verification
premixed, 3 plasmid kit; requires neon® transfection system or similar device
Kit contains all 4 genes required; cannot be purchased separately
price/unit $$ $ $$
available support resources standard protocol for fibroblast reprogramming
protocols for fibroblast reprogramming
protocols for fibroblast, cd34+, and pbmc reprogramming; faQs, webinar, publications
$—inexpensive $$—moderate $$$—expensive $$$$—exclusive
Other reprogramming toolsCells for reprogrammingThese cells can be reprogrammed to iPSCs using various reprogramming methods:
Fibroblasts
• Primary human dermal fibroblasts from either adult or neonatal samples
Keratinocytes• Primary human keratinocyte cells from either adult or
neonatal samples
CD34+ cells• Pooled human hematopoietic progenitor cells derived
from the umbilical cord blood of mixed donors
TaqMan® iPSC Sendai Detection Kitthese taqman® probes are used for detecting the presence and determining the levels of sendai virus and exogenous transcription factors (oct3/4, sox2, Klf4, and cmyc) delivered by the sendai virus from the cytotune®-ips sendai reprogramming Kit. the primers in the taqman® ipsc sendai detection Kit will not detect the corresponding endogenous factors.
Cat. No. A13640
Neon® Transfection SystemShockingly simple transfection in stem cellsthe neon® transfection system, a next-generation electroporation technology for highly efficient delivery (~80%) of nucleic acids such as episomal vectors into somatic cells.
Cat. No. MPK5000
10 life technologies—stem cell solutions
Validation is critical to induced pluripotent stem cell (iPSC) research. Whether you want to quickly confirm that cells are pluripotent, or need flexibility in antibody and dye choice, Molecular Probes® labeling products and detection assays are an ideal choice. And all can be visualized on the FLoid™ Cell Imaging Station. Backed by our extensive internal and external support resources including the Molecular Probes® Cell Imaging page on Facebook, and the trusted Molecular Probes® Handbook.
lifetechnologies.com/detectpsc
FLoid™ Cell Imaging StationCell imaging right at your benchtopconfidently detect and validate your esc and ipsc colonies with alkaline phosphatase live stain and cell surface antibodies on the affordable, user-friendly floid™ cell imaging station.
Cat. No. 4471136
Detection
Alkaline Phosphatase (AP) Live Stain shows robust staining in pluripotent stem cells. H9 human ESC grown on MEF feeders show specific staining of the pluripotent stem cell colonies.
H9 hESCs labeled with pluripotent surface antibodies: SSEA4 Alexa Fluor® 488 (green), TRA-1-81 Alexa Fluor® 594 (red), TRA-1-60 Alexa Fluor® 647 (purple).
Pluripotent stem cells (PSCs)
Alkaline Phosphatase Live StainQuick check for pluripotencyrapidly detect pluripotency in expanding escs and ipscs without compromising cell integrity with alkaline phosphatase live stain.
Cat. No. A14353
AntibodiesFlexibility and choicemix and match your choice of primary antibody and molecular probes® fluorophore for maximum flexibility in pluripotent stem cell imaging.
lifetechnologies.com/antibodies
lifetechnologies.com/stemcell 11
Analysis
Which detection method is right for you? Allows easy identification of
pluripotencyAllows easy identification of pluripotency without compromising cell integrity
Specific and flexible identification of PSCs; antibodies can be combined with different fluorophores for costaining
Product name ELF® 97 Endogenous Phosphatase Detection Kit
Alkaline Phosphatase Live Stain
Primary antibodies
cat. no. e6601 a14353 lifetechnologies.com/antibodies
specificity medium (stains stem and progenitor cells)
medium (stains stem and progenitor cells)
high (stains human escs and ipscs)
terminal assay Yes no no
reaction time stains pscs in 30 min stain pscs typically in 20 min or less
stain pscs in typically 90–120 min
color/label Yellow-green fluorescent precipitate under hoescht/dapi long pass filter
Green/fluorescein (fitc) unconjugated primary antibodies, may be detected with secondary conjugated to fluorophore of choice
floid™ protocols available no Yes Yes
unit size 500 µl vial sufficient for staining four 24-well plates, twelve 6-cm dishes, or four 10-cm dishes
50 µl vial sufficient for staining four 24-well plates, twelve 6-cm dishes, or four 10-cm dishes
100 µg vial sufficient for staining four 24-well plates, twelve 6-cm dishes, or four 10-cm dishes
relative price/unit $$ $ $$$$—inexpensive $$—moderate $$$—expensive $$$$—exclusive
Stem cell research requires cellular and molecular tools to confirm pluripotency of the stem cell population, or the success of a differentiation experiment. Whether analyzing proliferation, protein levels, gene expression, or epigenetic profiles, Life Technologies has the right instruments and products for your research. Go to www.lifetechnologies.com/stemcellanalysis
Selected Instruments
Ion Proton™ Sequencer Genome and exome sequencing on your benchtop
lifetechnologies.com/proton
QuantStudio™ 12K Flex Real-Time PCR System The all-in-one qPCR instrument
Delivering our highest throughput, flexibility, and scalability, the QuantStudio™ 12K Flex Real-Time PCR System features 5 interchangeable blocks to support 96-well, Fast 96-well, 384-well, TaqMan® Array Card, and OpenArray® formats.
lifetechnologies.com/quantstudio
Assays
TaqMan® Protein AssaysTaqMan® Protein Expression Assays are designed for accurate quantitation of protein expression using gold-standard TaqMan® 5’ nuclease chemistry with a workflow and sample quantity similar to our TaqMan® Gene Expression and MicroRNA (miRNA) Assays. As the protein expression results are obtained on the same analytical platform, these revolutionary assays enable direct correlation of mRNA and/or miRNA expression to protein expression.
lifetechnologies.com/taqmanprotein
12 life technologies—stem cell solutions
Mesenchymal stem cells (MSCs)Culture
MesenPRO RS™ MediumA reduced-serum medium for MSC culture containing 2% FBS mesenpro rs™ medium consistently improves expansion of mscs compared with classical media (dmem + 10% fbs) and maintains trilineage mesoderm differentiation potential.
Cat. No. 12746012
Mesenchymal stem cell research requires different culture conditions depending on your needs. Life Technologies provides the broadest selection of Gibco® cGMP compliant complete culture systems for mesenchymal stem cells (MSCs) and adipose-derived stem cells (ADSCs), some of which are free of animal-derived components. These media are designed to minimize adaptation time, maximize cell performance, and meet regulatory requirements. For more information, go to lifetechnologies.com/stemcell/msc
Which mesenchymal stem cell culture medium is right for you?The standard serum-based culture for MSCs and ADSCs Reduced serum medium for lower variability and
cost-savings with serum-cultureFully documented serum-free MSC culture reducing the burden in qualifying reagents, facilitating transition from the bench to the clinic
Serum-free, xeno-free MSC culture reducing the risk and variability associated with animal origin components
Product name MSC-Qualified FBS MesenPro RS™ Medium StemPro® MSC SFM CTS™ StemPro® MSC SFM XenoFree
cat. no. 12662-002, 12662-011, 12662-029 (usda), 12664-025 (australia origin), 12763-025 (us origin)
12746012 e071000 (new Zealand origin)
a1033201 a1067501
Recommended applications Multiple species; culture of cells sourced from bone marrow, adipose tissue, cord blood, umbilical cord, placenta
Multiple species; culture of cells sourced from bone marrow, adipose tissue, umbilical cord
Human; culture of cells sourced from bone marrow, adipose tissue, umbilical cord, hair follicle
Human; culture of cells sourced from bone marrow, adipose tissue, umbilical cord, and cord blood, as well as pericytes and fibroblasts
lot-to-lot variability high medium low low
components/unit 1 x 50 ml, 100 ml, or 500 ml 1 x 500 ml mesenpro rs™ basal medium 1 x 10 ml mesenpro rs™ Growth supplement
1 x 500 ml stempro® msc sfm basal medium 1 x 75 ml stempro® sfm supplement
1 x 500 ml stempro® msc sfm basal medium 1 x 5 ml stempro® msc sfm xenofree supplement
additional components required for complete medium
dmem low glucose Glutamax™-i medium
Glutamax™-i medium Glutamax™-i cts™, substrate (cellstart™ cts™ or attachment factor) Glutamax™-i cts™, substrate (cellstart™ cts™ or attachment factor)
supports growth at high cell density; less medium, surface area, and time
no no Yes Yes
relative price/500 ml of complete medium $ $ $$ $$$
stability of complete medium stable at 4°c for 2 weeks stable at 4°c for 15 days stable at 4°c for 4 weeks stable at 4°c for 2 weeks
specifically designed for cell therapy research
no no Yes Yes
$—inexpensive $$—moderate $$$—expensive $$$$—exclusive
Human MSCs from a 4-donor, passage 4 pool, expanded for 9 passages in StemPro® MSC SFM XenoFree on CELLstart™ CTS™ Substrate–coated flasks.
StemPro® MSC Serum Free Medium (SFM) CTS™
Serum-free medium for growth and expansion of MSCsstempro®msc sfm cts™ provides superior efficiency of human msc expansion at high cell densities, requiring less medium, surface area, and time compared with classical medium (dmem (low glucose) + 10% fbs) when used in conjunction with cellstart™ cts™ substrate.
Cat. No. A1033201
lifetechnologies.com/stemcell 13
Which mesenchymal stem cell culture medium is right for you?The standard serum-based culture for MSCs and ADSCs Reduced serum medium for lower variability and
cost-savings with serum-cultureFully documented serum-free MSC culture reducing the burden in qualifying reagents, facilitating transition from the bench to the clinic
Serum-free, xeno-free MSC culture reducing the risk and variability associated with animal origin components
Product name MSC-Qualified FBS MesenPro RS™ Medium StemPro® MSC SFM CTS™ StemPro® MSC SFM XenoFree
cat. no. 12662-002, 12662-011, 12662-029 (usda), 12664-025 (australia origin), 12763-025 (us origin)
12746012 e071000 (new Zealand origin)
a1033201 a1067501
Recommended applications Multiple species; culture of cells sourced from bone marrow, adipose tissue, cord blood, umbilical cord, placenta
Multiple species; culture of cells sourced from bone marrow, adipose tissue, umbilical cord
Human; culture of cells sourced from bone marrow, adipose tissue, umbilical cord, hair follicle
Human; culture of cells sourced from bone marrow, adipose tissue, umbilical cord, and cord blood, as well as pericytes and fibroblasts
lot-to-lot variability high medium low low
components/unit 1 x 50 ml, 100 ml, or 500 ml 1 x 500 ml mesenpro rs™ basal medium 1 x 10 ml mesenpro rs™ Growth supplement
1 x 500 ml stempro® msc sfm basal medium 1 x 75 ml stempro® sfm supplement
1 x 500 ml stempro® msc sfm basal medium 1 x 5 ml stempro® msc sfm xenofree supplement
additional components required for complete medium
dmem low glucose Glutamax™-i medium
Glutamax™-i medium Glutamax™-i cts™, substrate (cellstart™ cts™ or attachment factor) Glutamax™-i cts™, substrate (cellstart™ cts™ or attachment factor)
supports growth at high cell density; less medium, surface area, and time
no no Yes Yes
relative price/500 ml of complete medium $ $ $$ $$$
stability of complete medium stable at 4°c for 2 weeks stable at 4°c for 15 days stable at 4°c for 4 weeks stable at 4°c for 2 weeks
specifically designed for cell therapy research
no no Yes Yes
$—inexpensive $$—moderate $$$—expensive $$$$—exclusive
StemPro® MSC SFM XenoFreeEnabling serum-free and xeno-free expansion of human MSCsstempro® msc sfm xenofree offers a completely animal origin–free system when used in conjunction with cellstart™ cts™ substrate; thus, mscs and adscs are grown in a more physiologically similar environment that allows for more clinically relevant results.
Cat. No. A1067501
StemPro® LipoMAX™ SupplementDefined xeno-free lipid supplementstempro® lipomax™ supplement is a human-derived, lipoprotein-based cell culture supplement for the improved expansion of mscs and adscs.
Cat. No. A1085001
CELLstart™ CTS™ Substratexeno-free, fully defined cell culture substrate that contains components only of human origin. cellstart™ cts™ substrate enables attachment of human mscs and adscs.
Cat. No. A1014201
14 life technologies—stem cell solutions
Mesenchymal stem cells differentiated into fat cells and stained with Oil Red O, a fat-soluble dye.
Fluorescent visualization of StemPro® Rat Alk Phos Expressing Mesenchymal Stem Cells using ELF® 97 Endogenous Phosphatase Kit.
StemPro® Human Adipose-Derived Stem Cell (ADSC) Kithuman adscs isolated from human lipoaspirate tissue and cryopreserved from primary cultures. Kit contains human adscs and mesenpro rs™ medium.
Cat. No. R7788110
Mouse (C57BL⁄6) Mesenchymal Stem Cellsisolated from bone marrow of c57bl⁄6 mouse at ≤8 weeks after gestation.
Cat. No. S1502100
StemPro® Rat Alk Phos Expressing Mesenchymal Stem Cellsthese cells have a unique ability to track cells in transplantation and differentiation studies.
Cat. No. R7789120
Gibco® MSC differentiation kits Human MSC differentiation to the adipogenesis, chondrogenesis, and osteogenesis lineages.
Complement StemPro® MSC SFM, MesenPRO RS™ Medium, and MSC-qualified FBS cell expansion systems.
StemPro® Adipogenesis Differentiation Kitcomplete differentiation into fat cells.
Cat. No. A1007001
StemPro® Chondrogenesis Differentiation Kitcomplete differentiation into cartilage cells.
Cat. No. A1007101
StemPro® Osteogenesis Differentiation Kitcomplete differentiation into bone cells.
Cat. No. A1007201
Differentiation
Mesenchymal stem cells (MSCs)Culture
lifetechnologies.com/stemcell 15
A B C
D E
CD19 Pacific Blue™ fluorescence
CD14 Pacific Blue™ fluorescence HLA DR Pacific Blue™ fluorescence
CD45 Pacific Orange™ fluorescence FxCycle™ Violet fluorescence
CD73
Phy
coer
ythr
in fl
uore
scen
ce
CD73
Phy
coer
ythr
in fl
uore
scen
ce
Coun
t
CD90
FIT
C flu
ores
cenc
e
CD10
5 Pe
rCP-
Cy®
5.5
fluor
esce
nce
Immunophenotype data. Human mesenchymal stem cells collected at passage 7 were identified as: (A) 99.7% CD73+/0.29% CD19–; (B) 99.4% CD73+/0.22% CD45–; (C) 99.7% CD90+/0.09% CD34–; (D) 99.9% CD90+/0.07% CD14–; (E) 99.2% CD105+/0.76% HLA-DR–. Detection was by FACS using the Attune® Acoustic Focusing Cytometer, and the data demonstrate that cell expression remains as expected after 7 passages.
Detection
Detection of human mesenchymal stem cells using the Attune® Acoustic Focusing Cytometer For more information, go to lifetechnologies.com/attune
The International Society for Cellular Therapy (ISCT) has proposed a set of standards to define hMSCs for laboratory investigations and preclinical studies, including specific surface antigen expression in which ≥95% of the cells express the antigens recognized by CD105, CD73, and CD90, with the same cells lacking (≤2% positive) the antigens recognized by CD45, CD34, CD14 or CD11b, CD79a or CD19, and HLA-DR (Cytotherapy 8:315 (2006)).
16 life technologies—stem cell solutions
Neural stem cells (NSCs)Culture
DifferentiationB-27® Supplementsb-27® supplements are 50x liquid supplements for growth and long-term viability of hippocampal neurons. b-27® supplement is the most referenced supplement in neurobiology research, and is offered in both serum-free and xeno-free versions.lifetechnologies.com/b27
N-2 Supplementn-2 supplement is recommended for the growth and expression of neuroblastomas as well as postmitotic neurons in primary cultures from both the peripheral nervous system (pns) and the central nervous system (cns).
Cat. No. 17502048
These supplements are used in combination with a basal medium such as Neurobasal® Medium.
StemPro® NSC SFMserum-free growth and expansion of human neural stem cells (hnscs), with versatility to support both adherent and suspension nsc cultures.
Cat. No. A1050901
Neurobasal® and Neurobasal®-A mediaserum-free basal media for the long-term viability of hippocampal and other neurons of the central nervous system. neurobasal® medium is optimized for prenatal and embryonic neurons, while neurobasal®-a medium is best for growing postnatal and adult brain neurons.lifetechnologies.com/neurobasal
Natural Mouse Lamininsupports the growth of human or rat neural stem cells, as well as neural primary cells.
Cat. No. 23017015
Human Neural Stem Cells (H9-Derived)consistent high-purity cells with normal human karyotypes that can differentiate into neurons, oligodendrocytes, and astrocytes. consistent proliferation in adherent cell culture when used with stempro® nsc sfm.
Cat. No. N7800100
Rat Fetal Neural Stem Cellsthese cells can be expanded as adherent or neurosphere suspension cell cultures while more than 75% of nscs retain undifferentiated phenotype. also supports differentiation into neurons, oligodendrocytes, and astrocytes.
cat. no. n7744100
Human embryonic stem cells differentiated into neurons.
Gibco® media and supplements are the most widely cited reagents for use in neural stem cell culture. Choose between the flexibility of our B-27®, N-2, StemPro® Neural supplements, and Neurobasal® Medium, or ease of use with the StemPro® NSC SFM kit, specifically formulated for serum-free growth and expansion of human neural stem cells. For more information, go to lifetechnologies.com/stemcell/nsc
lifetechnologies.com/stemcell 17
Hematopoietic stem cells (HSCs)Culture
CD34+ cells reprogrammed into induced pluripotent stem cells using the
CytoTune®-iPS Sendai Reprogramming Kit.
Dynal® CD34 Progenitor Cell Selection Systemstem cells can be isolated directly from whole blood, cord blood, or bone using this positive magnetic isolation of human cd34+ progenitor stem cells with bead release.
Cat. No. 11301D
Hematopoietic stem cells expressing CD34 (CD34+ cells) are defined by their ability to replenish all blood cell types and to self-renew. We offer CD34+ media, cells, and selection tools specifically formulated to support the culture of these stem cells. For more information, go to lifetechnologies.com/stemcell/hsc
StemPro®-34 SFMserum-free medium specifically formulated to support the development of human hematopoietic stem cells (cd34+) from sources including bone marrow, peripheral blood, and neonatal cord blood. it is manufactured with components of animal, human, recombinant, or synthetic origin.
Cat. No. 10639011
StemPro® CD34+ Cell Kitpooled human hematopoietic progenitor cells (hpcs) derived from the umbilical cord blood of mixed donors. along with cord blood–derived cd34+ cells, this kit includes stempro®-34 sfm basal liquid medium and frozen stempro®-34 nutrient supplement to facilitate the immediate culturing of the stempro® cd34+ cells.
Cat. No. A14059
18 life technologies—stem cell solutions
Generating unblemished iPSCs for disease investigation
“If you alter the genetics of the cell type you’re working with by integrating viruses, you’re going to have an enormous amount of difficulty teasing out what is the backbone genetic contribution versus the altered cell type contribution. By being able to create cells that are intact and have no modifications to the genome, you can really feel secure that what you’re looking at now is the genetic contribution to the disease.
“We were of course thrilled to see that Life Technologies had made available a new product, the Sendai virus reprogramming product [CytoTune®-iPS Sendai Reprogramming Kit], which is as simple as the lentiviral reprogramming or retroviral reprogramming but has this major advantage that there are no reprogramming factors left. It’s fully nonintegrating.
“It makes use of this RNA virus, the Sendai virus, that’s nonreplicative. You infect the somatic cells just one time. Usually within 18 to 21 days we see the appearance of iPS cell colonies. After we’ve expanded those colonies and actually established a line, by the time they’ve been passaged between five to eight times there’s no presence of this RNA virus anymore. So the lines in essence have an unblemished genome and they have none of the reprogramming factors left. So you’ve really got the best possible material for doing the types of studies we’re interested in.”
Watch the video at lifetechnologies.com/ipscvideo
Chad Cowan, PhD, Assistant Professor
of Stem Cell and Regenerative Biology at
Harvard University
lifetechnologies.com/stemcell 19
Introducing LifeLab™ hands-on pluripotent stem cell workshops
Life Technologies is proud to announce the opening of LifeLab™ Stem Cell Research Centers in Carlsbad, CA, and Madison, WI. These centers provide customers with hands-on training for culturing and characterizing human embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs), as well as reprogramming techniques for the creation of iPSCs.
Find out about our global workshops at lifetechnologies.com/lifelab
For research use only. Not intended for any animal or human therapeutic or diagnostic use. ©2012 Life Technologies Corporation. All rights reserved. The trademarks mentioned herein are the property of Life Technologies Corporation or their respective owners. Accutase is a registered trademark of Innovative Cell Technologies. CytoTune is a registered trademark of DNAVEC Corporation. TaqMan® is a registered trademark of Roche Molecular Systems, Inc., used under permission and license. B-27 is a registered trademark of Southern Illinois University. Cy is a registered trademark of GE Healthcare. Facebook is a trademark of Facebook, Inc. CO25406 0512
For more information, go to lifetechnologies.com/stemcell
publications—protocols—videos—access to experts