Enrichment and purging of human embryonic stem cells by detection of cell surface antigens using the monoclonal antibodies TG30 and GCTM-2

Journal of Visualized Experiments : JoVE
Juan Carlos PolancoAndrew L Laslett

Abstract

Human embryonic stem cells (hESC) can self-renew indefinitely in vitro, and with the appropriate cues can be induced to differentiate into potentially all somatic cell lineages. Differentiated hESC derivatives can potentially be used in transplantation therapies to treat a variety of cell-degenerative diseases. However, hESC differentiation protocols usually yield a mixture of differentiated target and off-target cell types as well as residual undifferentiated cells. For the translation of differentiated hESC-derivatives from the laboratory to the clinic, it is important to be able to discriminate between undifferentiated (pluripotent) and differentiated cells, and generate methods to separate these populations. Safe application of hESC-derived somatic cell types can only be accomplished with pluripotent stem cell-free populations, as residual hESCs could induce tumors known as teratomas following transplantation. Towards this end, here we describe a methodology to detect pluripotency associated cell surface antigens with the monoclonal antibodies TG30 (CD9) and GCTM-2 via fluorescence activated cell sorting (FACS) for the identification of pluripotent TG30(Hi)-GCTM-2(Hi) hESCs using positive selection. Using negative selection...Continue Reading

References

Mar 6, 2007·BMC Developmental Biology·Andrew L LaslettMartin F Pera
Jun 19, 2007·Nature Biotechnology·UNKNOWN International Stem Cell InitiativeWeidong Zhang
Jul 11, 2009·Nature Biotechnology·Kyoko MiuraShinya Yamanaka
Jun 11, 2010·Nature·Martin F Pera, Patrick P L Tam
Jun 25, 2010·Current Protocols in Stem Cell Biology·Qi ZhouAndrew L Laslett
Jan 18, 2012·Expert Opinion on Drug Metabolism & Toxicology·Francesca PistollatoGlyn Stacey
May 24, 2012·Toxicology and Applied Pharmacology·Michaela E SharpeAnnamaria Rossi

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Methods Mentioned

BETA
fluorescence activated cell sorting
enzymatic dissociation
FACS

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