Development of genetic quality tests for good manufacturing practice-compliant induced pluripotent stem cells and their derivatives

Purpose : We investigated RNA-seq-based broad-range genetic quality tests on GMP-compliant human leucocyte antigen (HLA)-homozygous hiPSCs and their derivatives under postdistribution conditions to investigate whether sequencing data could provide a basis for future quality control.Methods : Transcriptome profiling of the differentiated cell lines in three lineages, including neuronal cells, cardiomyocytes and hepatocytes, at early and late passages, respectively, were carried out using RNA sequencing.Results : We performed SNV, indel, CNV, ekaryotyping, HLA genotyping and transcriptomic analyses in which differentiated cells were compared with hiPSCs using RNA-seq data from undifferentiated and differentiated cultures returned from multiple centres. In particular, we evaluated whether prolonged culture conditions have the potential to give rise to genomic and phenotypic changes in the CMC3 hiPSC line, a candidate line for manufacturing cell therapies.Conclusion : RNA-seq analysis appears to offer an informative genetic quality testing approach for cell types and allows the early screening of candidate hiPSC seed stocks for clinical use by facilitating safety and potential risk evaluation