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Ask the Expert – Options for Identification and Characterization of Pluripotent Stem Cells
The vast advances in technologies for the efficient generation of footprint-free induced pluripotent stem cells (iPSC) have led to the creation of several iPSC lines from varying sources, genetic backgrounds, and derivations in different medias and growth conditions, thus necessitating thorough characterization of the resulting cell lines.
One critical step in establishing iPSC lines involves the early identification of true iPSC clones and their subsequent characterization to ensure functional pluripotency. Various methods of characterization, ranging from visual morphological observation to the use of differentially expressed biomarkers are utilized for the initial identification of pluripotent cells. Dyes such as Alkaline Phosphatase Live Stain enable the early detection of emerging iPSC colonies that can be used in combination with morphological assessment to pick the right iPSC clone for further expansion. Established clones are further subjected to a combination of in vitro and in vivo cellular analysis to confirm functional pluripotency based on expression of self-renewal markers and trilineage differentiation potential. While such traditional methods have been successfully used, there is a need for a uniform standardized method for comprehensive characterization. Recently, the TaqMan® hPSC Scorecard™ Panel, a real-time PCR gene expression assay, provides a rapid molecular method to generate quantitative transcriptome data for the confirmation of functional pluripotency.
Join Uma Lakshmipathy, Principal Scientist at Thermo Fisher Scientific, to explore various options for identification and characterization of pluripotent stem cells.
Dr Uma Lakshmipathy has been involved in the field of stem cells for nearly a decade. Her doctoral degree in Molecular Biophysics and subsequent postdoctoral experience in DNA repair brought new perspective and led her to the area of stem cell research with focus on developing ex vivo gene repair systems. As a junior faculty at the Stem Cell Institute, University of Minnesota, she identified efficient gene delivery methods into stem cells to enable repair of adult stem cells from monogene disorders. She moved to Invitrogen, Life Technologies, in 2005 and was involved in the development of novel technology platforms for creating labeled stem cells. Her current research interests at Life Technologies, now a part of Thermo Fisher Scientific, are regulation of stem cell maintenance, development of technologies for generation, identification, characterization of pluripotent stem cells, and modification their derivatives.
Don’t miss this chance to have your identification and characterization questions answered, session starts Monday.