A panel of tests to standardize the characterization of human embryonic stem cells

Abstract

Human embryonic stem cells offer a renewable source of a wide range of cell types for use in research and cell-based therapies. Characterizing these cells provides important information about their current state and affords relevant details for subsequent downstream manipulation. Prior to considering therapeutic applications, it is crucial that the cells are surveyed at a genetic and proteomic level during the extensive propagation, expansion and differentiation. Hence, a set of characterization tests to measure stem cell stability and identity – genomic, epigenomic and mitochondrial markers, as well as functional measures of utility, need to be developed. Thus, we outline a plan of standard assays that can be afforded by multiple laboratories to unambiguously test the quality of human embryonic stem cells. In this manuscript, we describe a comprehensive characterization of ReliCell®hES1, the only human embryonic stem cell line reported from the Indian subcontinent. Our study employs gene expression analysis using quantitative reverse transcription-polymerase chain reaction and microarray, mitochondrial DNA sequencing, microRNA analysis, immunophenotyping and teratoma formation, in addition to demonstrating its capacity to propagate under feeder-free conditions.