Reliable multiplex generation of pooled induced pluripotent stem cells.
CP: Stem cell
bioinformatics
deconvolution
differentiation
flow cytometry
genomics
iPSCs
multiplex
organoids
reprogramming
sequencing
Journal
Cell reports methods
ISSN: 2667-2375
Titre abrégé: Cell Rep Methods
Pays: United States
ID NLM: 9918227360606676
Informations de publication
Date de publication:
25 09 2023
25 09 2023
Historique:
received:
28
06
2022
revised:
23
06
2023
accepted:
04
08
2023
medline:
28
9
2023
pubmed:
27
9
2023
entrez:
26
9
2023
Statut:
ppublish
Résumé
Reprogramming 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.
Identifiants
pubmed: 37751688
pii: S2667-2375(23)00214-X
doi: 10.1016/j.crmeth.2023.100570
pmc: PMC10545906
pii:
doi:
Types de publication
Journal Article
Research Support, N.I.H., Extramural
Langues
eng
Sous-ensembles de citation
IM
Pagination
100570Subventions
Organisme : NIDCD NIH HHS
ID : R21 DC018092
Pays : United States
Organisme : NIA NIH HHS
ID : U01 AG061835
Pays : United States
Informations de copyright
Copyright © 2023 The Authors. Published by Elsevier Inc. All rights reserved.
Déclaration de conflit d'intérêts
Declaration of interests G.M.C. holds leadership positions in many companies related to DNA sequencing technologies. A full list of these companies is available at http://arep.med.harvard.edu/gmc/tech.html.
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