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

Abstract：

:Although human induced pluripotent stem cell (hiPSC) lines are karyotypically normal, they retain the potential for mutation in the genome. Accordingly, intensive and relevant quality controls for clinical-grade hiPSCs remain imperative. As a conceptual approach, we performed 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. We found differences in the degree of single-nucleotide polymorphism (SNP) occurring in cells cultured at three collaborating institutes. However, the cells cultured at each centre showed similar trends, in which more SNPs occurred in late-passage hiPSCs than in early-passage hiPSCs after differentiation. In eSNP karyotyping analysis, none of the predicted copy number variations (CNVs) were identified, which confirmed the results of SNP chip-based CNV analysis. HLA genotyping analysis revealed that each cell line was homozygous for HLA-A, HLA-B, and DRB1 and heterozygous for HLA-DPB type. Gene expression profiling showed a similar differentiation ability of early- and late-passage hiPSCs into cardiomyocyte-like, hepatic-like, and neuronal cell types. However, time-course analysis identified five clusters showing different patterns of gene expression, which were mainly related to the immune response. In conclusion, RNA-seq analysis appears to offer an informative genetic quality testing approach for such cell types and allows the early screening of candidate hiPSC seed stocks for clinical use by facilitating safety and potential risk evaluation.

journal_name

Sci Rep

journal_title

Scientific reports

authors

Jo HY,Han HW,Jung I,Ju JH,Park SJ,Moon S,Geum D,Kim H,Park HJ,Kim S,Stacey GN,Koo SK,Park MH,Kim JH

doi

10.1038/s41598-020-60466-9

subject

Has Abstract

pub_date

2020-03-03 00:00:00

pages

3939

issue

1

issn

2045-2322

pii

10.1038/s41598-020-60466-9

journal_volume

10

pub_type

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