Reliable multiplex generation of pooled induced pluripotent stem cells for genetic testing

Abstract

Inducing somatic cells into pluripotent stem cells (iPSCs) provides an excellent model for studying systems in-vitro. Understanding the impact of individual donor genetic backgrounds on reprogramming ability would allow researchers to harness these genetic differences and increase the efficiency of the reprogramming process. To better understand the genetic basis of reprogramming cells into iPSCs, we present induction of Pluripotency from Pooled Cells (iPPC) - an efficient, scalable, and reliable reprogramming procedure. Using our deconvolution algorithm that employs low-coverage pooled sequencing and single nucleotide polymorphisms (SNPs), we estimate individual donor proportions of cell lines within large cohorts. With iPPC, we concurrently reprogrammed over one hundred donor LCLs into iPSCs and found strong correlations of individual donors' reprogramming ability across multiple experiments. We note that individual donors' reprogramming ability remains consistent across both same-day replicates and multiple experimental runs, and that the expression of certain immunoglobulin precursor genes (IGLV10-54, IGLV3-9, IGLV1-17, IGLV1-6, and IGLV3-1) may impact reprogramming ability. Our process enables a multiplex framework to study the reprogramming ability of different donor cells into iPSCs and also provides a reliable method along with a pooled library of donor iPSCs for downstream research and investigation of other in-vitro phenotypes.

Development of a high-throughput, quantitative platform using human cerebral organoids to study virus-induced neuroinflammation in Alzheimer’s disease

“…The control donor hiPSC line used in our study (PGP1 iPSC) was a kind gift from Professor George Church's lab, and we had previously characterized and sequenced the line 34,88 . We performed flow cytometry (MACSquant VYB) to confirm that >90% of the PGP1 hiPSCs are positive for TRA-1-60 (Novus Biologicals NB100-730F488).…”