Smullen, MollyOlson, Meagan NReichert, Julia MDawes, PepperMurray, Liam FBaer, Christina EWang, QiReadhead, BenjaminChurch, George MLim, Elaine TChan, Yingleong2023-10-232023-10-232023-08-31Smullen M, Olson MN, Reichert JM, Dawes P, Murray LF, Baer CE, Wang Q, Readhead B, Church GM, Lim ET, Chan Y. Reliable multiplex generation of pooled induced pluripotent stem cells. Cell Rep Methods. 2023 Sep 25;3(9):100570. doi: 10.1016/j.crmeth.2023.100570. Epub 2023 Aug 31. PMID: 37751688; PMCID: PMC10545906.2667-237510.1016/j.crmeth.2023.10057037751688https://hdl.handle.net/20.500.14038/52637Reprogramming somatic cells into pluripotent stem cells (iPSCs) enables the study of systems in vitro. To increase the throughput of reprogramming, we present induction of pluripotency from pooled cells (iPPC)-an efficient, scalable, and reliable reprogramming procedure. Using our deconvolution algorithm that employs pooled sequencing of single-nucleotide polymorphisms (SNPs), we accurately estimated individual donor proportions of the pooled iPSCs. With iPPC, we concurrently reprogrammed over one hundred donor lymphoblastoid cell lines (LCLs) into iPSCs and found strong correlations of individual donors' reprogramming ability across multiple experiments. Individual donors' reprogramming ability remains consistent across both same-day replicates and multiple experimental runs, and the expression of certain immunoglobulin precursor genes may impact reprogramming ability. The pooled iPSCs were also able to differentiate into cerebral organoids. Our procedure enables a multiplex framework of using pooled libraries of donor iPSCs for downstream research and investigation of in vitro phenotypes.enCopyright 2023 The Authors. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/)Attribution-NonCommercial-NoDerivatives 4.0 Internationalhttp://creativecommons.org/licenses/by-nc-nd/4.0/CP: Stem cellbioinformaticsdeconvolutiondifferentiationflow cytometrygenomicsiPSCsmultiplexorganoidsreprogrammingsequencingJournal ArticleCell reports methods