Human Pluripotent Stem Cells for High-Throughput Drug Screening and Characterization of Small Molecules.
Cell viability
Dose–response curves
Embryonic stem cells
High-throughput screening
Induced pluripotent stem cells
Robotic cell culture
Small molecules
Toxicity
Journal
Methods in molecular biology (Clifton, N.J.)
ISSN: 1940-6029
Titre abrégé: Methods Mol Biol
Pays: United States
ID NLM: 9214969
Informations de publication
Date de publication:
2022
2022
Historique:
pubmed:
16
6
2021
medline:
24
6
2022
entrez:
15
6
2021
Statut:
ppublish
Résumé
Human pluripotent stem cells (hPSCs), such as induced pluripotent stem cells (iPSCs), hold great promise for drug discovery, toxicology studies, and regenerative medicine. Here, we describe standardized protocols and experimental procedures that combine automated cell culture for scalable production of hPSCs with quantitative high-throughput screening (qHTS) in miniaturized 384-well plates. As a proof of principle, we established dose-response assessments and determined optimal concentrations of 12 small molecule compounds that are commonly used in the stem cell field. Multi-parametric analysis of readouts from diverse assays including cell viability, mitochondrial membrane potential, plasma membrane integrity, and ATP production was used to distinguish normal biological responses from cellular stress induced by small molecule treatment. Collectively, the establishment of integrated workflows for cell manufacturing, qHTS, high-content imaging, and data analysis provides an end-to-end platform for industrial-scale projects and should leverage the drug discovery process using hPSC-derived cell types.
Identifiants
pubmed: 34128205
doi: 10.1007/7651_2021_394
pmc: PMC9520585
mid: NIHMS1831765
doi:
Types de publication
Journal Article
Research Support, N.I.H., Extramural
Research Support, N.I.H., Intramural
Langues
eng
Sous-ensembles de citation
IM
Pagination
811-827Subventions
Organisme : Intramural NIH HHS
ID : Z99 TR999999
Pays : United States
Commentaires et corrections
Type : ErratumIn
Informations de copyright
© 2021. Springer Science+Business Media, LLC.
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