Stem cell research solutions and support

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Stem cell research can be time-consuming. Finding support shouldn t be. 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. Go to lifetechnologies.com/pscresources to check out protocols, FAQs, publications, scientific posters, videos, and upcoming events. Watch the experts Hands-on training Technical resources Ask the experts 2 Life Technologies Stem cell solutions

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 lifetechnologies.com/stemcell 3

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 Data image for KnockOut SR XenoFree CTS Kit provided by Dr. Hidenori Akutsu, National Center for Child Health and Development, Tokyo, Japan. KnockOut Serum Replacement (KSR) Well-referenced, proven standard KnockOut Serum Replacement and basal media deliver confidence, with over 2,400 peer-reviewed publications. Cat. No. 10828010, 10828028 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 Product name KnockOut Serum Replacement StemPro hesc SFM Cat. No. 10828010, 10828028 A1000701 Recommended applications Multiple species; feeder culture; reprogramming on feeders; differentiation of PSCs Human; feeder-free culture Lot-to-lot variability Medium 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 Components/unit 1 x 100 ml or 500 ml 1 x 100 ml StemPro hesc Supplement 1 x 500 ml DMEM/F12 with GlutaMAX Medium 1 x 50 ml 25% BSA Additional components required for complete medium Price/500 ml of complete medium DMEM/F12 (1X) with GlutaMAX medium, NEAA, bfgf; 2-mercaptoethanol; for a cost-effective option, choose the KnockOut ESC/iPSC Media Kit $$ individually $ for kit bfgf and 2-mercaptoethanol $$ Stability of complete medium Stable at 4 C for 2 weeks Stable at 4 C for 2 weeks Specifically designed for cell therapy research Available support resources No Protocols for culturing PSCs, publications, hands-on training, product finder No Protocols for culturing PSCs $ inexpensive $$ moderate $$$ expensive $$$$ exclusive 4 Life Technologies Stem cell solutions

StemPro hesc SFM Flexible feeder-free culture StemPro hesc SFM optimizes bfgf concentration and maintains the differentiation capability of PSCs. Cat. No. A1000701 Essential 8 Medium Feeder-free culture for all Essential 8 Medium contains only the necessary components for optimal PSC culture for every step of your research. Cat. No. A14666SA KnockOut SR XenoFree CTS Kit Traceability for cell therapy research The KnockOut SR XenoFree CTS Kit reduces the burden in qualifying reagents, facilitating transition from the bench to the clinic. Cat. No. A1448801, A1448701 Optimal feeder-free culture of all human ESC and ipsc lines with minimal lot-to-lot variability Essential 8 Medium A14666SA Human; feeder-free, xeno-free culture; feeder-free reprogramming Low Compact colonies with smooth edges; high nuclear to cytoplasmic ratio 1 x 10 ml Essential 8 Supplement 1 x 500 ml DMEM/F-12 (HAM) 1:1 No additional components required Fully documented xeno-free PSC culture reducing the burden in qualifying reagents, facilitating transition, from the bench to the clinic KnockOut CTS XenoFree ESC/iPSC Kit A1448801, A1448701 Human; feeder-based and feeder-free xeno-free culture for cell therapy research Medium Compact colonies with smooth edges; some colonies will have undefined borders with spikey edges 1 x 100 ml KnockOut SR XenoFree CTS 1 x 500 ml KnockOut DMEM CTS 1 x 5 ml KnockOut SR GF Cocktail CTS bfgf CTS and 2-mercaptoethanol $ $$$$ Stable at 4 C for 2 weeks No Protocols for culturing PSCs, FAQs, hands-on training Stable at 4 C for 2 weeks Yes Protocols for culturing PSCs, FAQs, product documentation lifetechnologies.com/stemcell 5

Pluripotent stem cells (PSCs) Complementary culture products 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 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 6 Life Technologies Stem cell solutions

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 feederfree cultures Standard reagent for clump passaging 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 Product name Dispase Collagenase IV Accutase UltraPure 0.5M EDTA Optimal for clinical research applications that require human origin materials only Quick and easy manual passaging tool for feeder and feeder-free cultures 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 Source origin Bacillus polymyxa Clostridium histolyticum Dissociation to single cells Marine Passaging in clumps Chemical (animal origin free (AOF)) Passaging in clumps or single-cell passaging Recombinant enzyme (AOF) Mechanical passaging in clumps 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 Inactivation method 2 weeks 2 weeks 2 years 2 years 2 years NA 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) Unit size 5 g 1 g 100 ml 4 x 100 ml 100 ml Box of 10 units Optimized concentration for use Number of plates passaged/unit Relative price/60 mm plate 2 mg/ml 1 mg/ml for feederdependent, 10 mg/ ml for feeder-free 1,250 x 60 mm plates 500 x 60 mm plates at 1 mg/ml, 50 x 60 mm plates at 10 mg/ml 1X solution 0.5 mm 1X solution NA 50 x 60 mm plates 50,000 x 60 mm plates (100 ml at 1,000x dilution) NA NA 50 x 60 mm plates 10 plates $ $ $$ $ $$ $$$ $ inexpensive $$ moderate $$$ expensive $$$$ exclusive An ipsc colony cut into evenly sized pieces using the StemPro EZPassage Disposable Stem Cell Passaging Tool. lifetechnologies.com/stemcell 7

Pluripotent stem cells (PSCs) Tools for reprogramming Selecting 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 A. CytoTune -ips Sendai Reprograming Kit B. Lentivirus C. CytoTune -ips Sendai Reprogramming Kit D. mrna (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 postreprogramming, ~130 ipsc colonies were observed from CytoTune reprogramming, while 11 ipsc colonies were observed from mrna reprogramming. ipsc colonies generated using Episomal ipsc Reprogramming Vectors Express TRA-1-81 (green) at Day 24. Lentiviral ipsc reprogramming particles Well-referenced, flexible technology Choose from eight different high-titer lentiviral ipsc reprogramming particles for your custom ipsc experiments. lifetechnologies.com/lenti 8 Life Technologies Stem cell solutions Episomal ipsc Reprogramming Vectors Safety with confidence Create reproducible ipscs from normal and diseased cell types with Invitrogen episomal vectors. CytoTune -ips Sendai Reprogramming Kit High efficiency for difficult cell types Reprogram challenging cell types with the easy-to-use, nonintegrating CytoTune -ips Sendai Reprogramming Kit.

Which reprogramming technology is right for you? Well-referenced technology for ipsc generation from fibroblasts Product name Lentiviral ipsc reprogramming particles Cat. No. A1357101, A1357201, A1357301, A1357401, A1357501, A1357601, A1357701, A1357801 Proven technology for viralfree ipsc generation from normal and diseased cell types Episomal ipsc Reprogramming Vectors A14703 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 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 Price/unit $$ $ $$ Available support resources $ inexpensive $$ moderate $$$ expensive $$$$ exclusive Standard protocol for fibroblast reprogramming Protocols for fibroblast reprogramming Efficient generation of integration-free ipscs from multiple somatic cell types CytoTune -ips Sendai Reprogramming Kit A1378001, A1378002 One application, minimal QC of putative colonies required Kit contains all 4 genes required; cannot be purchased separately Protocols for fibroblast, CD34+, and PBMC reprogramming; FAQs, webinar, publications Other reprogramming tools Cells for reprogramming These 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 Kit These 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 System Shockingly simple transfection in stem cells The 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 lifetechnologies.com/stemcell 9

Pluripotent stem cells (PSCs) Detection 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 Station Cell imaging right at your benchtop Confidently detect and validate your ESC and ipsc colonies with Alkaline Phosphatase Live Stain and cell surface antibodies on the affordable, userfriendly FLoid Cell Imaging Station. Cat. No. 4471136 Alkaline Phosphatase Live Stain Quick check for pluripotency Rapidly detect pluripotency in expanding ESCs and ipscs without compromising cell integrity with Alkaline Phosphatase Live Stain. Cat. No. A14353 Antibodies Flexibility and choice Mix and match your choice of primary antibody and Molecular Probes fluorophore for maximum flexibility in pluripotent stem cell imaging. lifetechnologies.com/antibodies 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). 10 Life Technologies Stem cell solutions

Which detection method is right for you? Product name Allows easy identification of pluripotency ELF 97 Endogenous Phosphatase Detection Kit Allows easy identification of pluripotency without compromising cell integrity Alkaline Phosphatase Live Stain Specific and flexible identification of PSCs; antibodies can be combined with different fluorophores for costaining 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 Color/label Yellow-green fluorescent precipitate under Hoescht/DAPI long pass filter Green/fluorescein (FITC) 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 Relative price/unit $$ $ $$$ $ inexpensive $$ moderate $$$ expensive $$$$ exclusive Stain PSCs in typically 90 120 min Unconjugated primary antibodies, may be detected with secondary conjugated to fluorophore of choice 100 µg vial sufficient for staining four 24-well plates, twelve 6-cm dishes, or four 10-cm dishes Analysis 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 Assays TaqMan 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 lifetechnologies.com/stemcell 11

Mesenchymal stem cells (MSCs) Culture 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 Human MSCs from a 4-donor, passage 4 pool, expanded for 9 passages in StemPro MSC SFM XenoFree on CELLstart CTS Substrate coated flasks. MesenPRO RS Medium A 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 StemPro MSC Serum Free Medium (SFM) CTS Serum-free medium for growth and expansion of MSCs StemPro 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 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-culture Product name MSC-Qualified FBS MesenPro RS Medium Cat. No. 12662-002, 12662-011, 12662-029 (USDA), 12664-025 (Australia origin), 12763-025 (US origin) Recommended applications Multiple species; culture of cells sourced from bone marrow, adipose tissue, cord blood, umbilical cord, placenta 12746012 E071000 (New Zealand origin) Multiple species; culture of cells sourced from bone marrow, adipose tissue, umbilical cord Lot-to-lot variability High Medium 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 Additional components required for complete DMEM low glucose GlutaMAX -I Medium medium GlutaMAX -I Medium Supports growth at high cell density; less No No medium, surface area, and time 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 Specifically designed for cell therapy research No No $ inexpensive $$ moderate $$$ expensive $$$$ exclusive 12 Life Technologies Stem cell solutions

StemPro MSC SFM XenoFree Enabling serum-free and xenofree expansion of human MSCs StemPro 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 Supplement Defined xeno-free lipid supplement StemPro LipoMAX Supplement is a human-derived, lipoprotein-based cell culture supplement for the improved expansion of MSCs and ADSCs. Cat. No. A1085001 CELLstart CTS Substrate Xeno-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 Fully documented serum-free MSC culture reducing the burden in qualifying reagents, facilitating transition from the bench to the clinic StemPro MSC SFM CTS A1033201 Serum-free, xeno-free MSC culture reducing the risk and variability associated with animal origin components StemPro MSC SFM XenoFree A1067501 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 Low 1 x 500 ml StemPro MSC SFM Basal Medium 1 x 75 ml StemPro SFM Supplement GlutaMAX -I CTS, substrate (CELLstart CTS or Attachment Factor) Low 1 x 500 ml StemPro MSC SFM Basal Medium 1 x 5 ml StemPro MSC SFM XenoFree Supplement GlutaMAX -I CTS, substrate (CELLstart CTS or Attachment Factor) Yes Yes $$ $$$ Stable at 4 C for 4 weeks Stable at 4 C for 2 weeks Yes Yes lifetechnologies.com/stemcell 13

Mesenchymal stem cells (MSCs) Culture StemPro Human Adipose-Derived Stem Cell (ADSC) Kit Human 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 Cells Isolated from bone marrow of C57BL 6 mouse at 8 weeks after gestation. Cat. No. S1502100 StemPro Rat Alk Phos Expressing Mesenchymal Stem Cells These cells have a unique ability to track cells in transplantation and differentiation studies. Cat. No. R7789120 Fluorescent visualization of StemPro Rat Alk Phos Expressing Mesenchymal Stem Cells using ELF 97 Endogenous Phosphatase Kit. Differentiation 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 Kit Complete differentiation into fat cells. Cat. No. A1007001 StemPro Chondrogenesis Differentiation Kit Complete differentiation into cartilage cells. Cat. No. A1007101 StemPro Osteogenesis Differentiation Kit Complete differentiation into bone cells. Cat. No. A1007201 Mesenchymal stem cells differentiated into fat cells and stained with Oil Red O, a fat-soluble dye. 14 Life Technologies Stem cell solutions

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)). A B C CD73 Phycoerythrin fluorescence CD73 Phycoerythrin fluorescence Count D CD19 Pacific Blue fluorescence E CD45 Pacific Orange fluorescence FxCycle Violet fluorescence CD90 FITC fluorescence CD105 PerCP-Cy 5.5 fluorescence CD14 Pacific Blue fluorescence HLA DR Pacific Blue fluorescence 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. lifetechnologies.com/stemcell 15

Neural stem cells (NSCs) Culture 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 StemPro NSC SFM Serum-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 media Serum-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 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 Cells These 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 Natural Mouse Laminin Supports the growth of human or rat neural stem cells, as well as neural primary cells. Cat. No. 23017015 Differentiation B-27 Supplements B-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 Supplement N-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. Human embryonic stem cells differentiated into neurons. 16 Life Technologies Stem cell solutions

Hematopoietic stem cells (HSCs) Culture Hematopoietic stem cells expressing CD34 (CD34+ cells) are defined by their ability to replenish all blood cell types and to selfrenew. 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 SFM Serum-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 CD34+ cells reprogrammed into induced pluripotent stem cells using the CytoTune -ips Sendai Reprogramming Kit. StemPro CD34+ Cell Kit Pooled 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 Dynal CD34 Progenitor Cell Selection System Stem 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 lifetechnologies.com/stemcell 17

Generating unblemished ipscs for disease investigation Chad Cowan, PhD, Assistant Professor of Stem Cell and Regenerative Biology at Harvard University 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 18 Life Technologies Stem cell solutions

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 lifetechnologies.com/stemcell 19

publications protocols videos access to experts For more information, go to lifetechnologies.com/stemcell 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