The revolutionary technology of converting adult somatic cells to pluripotent stem cells has led to the rapid progress of efficient, footprint-free methods for generation of induced pluripotent stem cells (iPSC). The resulting iPSC derived from diverse patient sources using different methods and conditions has created a challenge for rapid and comprehensive characterization. Traditional characterization methods are based on biomarker expression that utilizes a combination of in vitro and in vivo cellular analysis to confirm pluripotency and trilineage differentiation potential. Such methods are subjective and not amenable to high throughput confirmation.
TaqMan® hPSC Scorecard™ Panel is a comprehensive gene expression real-time PCR assay that can be utilized for rapid generation of quantitative transcriptome data. High density and medium density gene expression analyses were used to identify the optimal genes that define the pluripotent state and detect early stages of differentiation into cell types representative of the three germ layers. The resulting TaqMan® Human Pluripotent Stem Cell Scorecard consists of 93 assays comprising a combination of controls, pluripotent and lineage specific genes. Resulting expression data is analyzed using cloud-based analysis software that compares the expression pattern against a reference standard generated using multiple functionally validated ESC and iPSC lines. This system was successfully used to test several ESC and iPSC lines to confirm their pluripotency and determine their trilineage differentiation potential.