Improvements in induced pluripotent stem cell (iPSC) reprogramming technologies have led to the generation of patient-derived stem cells from various sources and conditions, creating valuable tools in drug discovery and future cell therapies. The steep challenge of characterizing these resulting iPSCs is minimally addressed by current methods that rely on a combination of in vitro and in vivo cellular methods. Molecular analysis methods offer an appealing solution for rapid, quantitative, and comprehensive characterization.

We have earlier reported the development of an hPSC Scorecard™ Panel comprising a 94-gene TaqMan® panel. The accompanying cloud-based analysis software computes the signature for self-renewal and lineage markers for test samples and compares the data against a pluripotent reference standard to generate scores. Over two hundred samples were analyzed using the Scorecard™ to determine pluripotency along several stages of the iPSC workflow. Established clones were allowed to differentiate spontaneously towards embryoid bodies to assess trilineage differentiation potential. Established clones were further used for directed differentiation towards specific lineages and the optimal combination of cytokines and length of differentiation was determined using Scorecard™. To further simplify the process and minimize sample size, methods were developed for the direct use of cell lysate in the assay without compromising the quality of the results.

These results collectively demonstrate the simplicity, ease and consistency of this method to predict functionality, thus offering a much needed uniform standardization and qualification of pluripotent cell lines.