EBiSC best practice: How to ensure optimal generation, qualification, and distribution of iPSC lines

Abstract

Disease-relevant human induced pluripotent stem cells (iPSCs) are generated worldwide for research purposes; however, without robust and practical ethical, legal, and quality standards, there is a high risk that their true potential will not be realized. Best practices for tissue procurement, iPSC reprogramming, day-to-day cultivation, quality control, and data management aligned with an ethical and legal framework must be included into daily operations to ensure their promise is maximized. Here we discuss key learning experiences from 7 years of operating the European Bank for induced Pluripotent Stem Cells (EBiSC) and recommend how to incorporate solutions into a daily management framework.

Homozygous might be hemizygous: CRISPR/Cas9 editing in iPSCs results in detrimental on-target defects that escape standard quality controls

“…A combination of simple-to-execute assays including allele copy number quantitative genotyping PCR (qgPCR) and assessment of heterozygosity through sequencing of nearby heterozygous single nucleotide polymorphisms (SNPs) successfully identified all lines that had appeared to be correctly edited, but were, in fact, defective. Our findings are in accordance with recent reports (Weisheit et al, 2020(Weisheit et al, , 2021 and further highlight the need for standardized quality control (QC) of CRISPR/Cas9-edited iPSC lines (for an overview of QC practices for iPSCs, see Steeg et al, 2021).…”

Generation of transgene-free human induced pluripotent stem cells from erythroblasts in feeder-free conditions

“…Prior to downstream use, hiPSCs need to pass several quality controls (QCs) ( Figure 4 ). These controls are part of the good practices for hiPSC research as described by the EBiSC ( Steeg et al., 2021 ; Sullivan et al., 2018 ). These QCs are absolutely necessary to ensure of the good quality of the hiPSCs.…”

Creation of GMP-Compliant iPSCs From Banked Umbilical Cord Blood

“…The BMDI CBB has a highly successful donor follow-up program, with CB donors re-consenting for their banked CB to be used to create iPSCs for potential therapeutic and commercial use (manuscript submitted). All aspects encompassing consent, ethics and a robust quality management framework are in place in accordance with worlds’ best practice ( Steeg et al, 2021 ) to adopt the protocol described here for creation of GMP-grade CB-derived iPSC.…”