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  2. Human Pluripotent Stem Cells for High-Throughput Drug Screening and Characterization of Small Molecules

Human Pluripotent Stem Cells for High-Throughput Drug Screening and Characterization of Small Molecules

  • Methods Mol Biol. 2022;2454:811-827. doi: 10.1007/7651_2021_394.
Seungmi Ryu 1 Pei-Hsuan Chu 1 Claire Malley 1 John Braisted 1 Pinar Ormanoglu 1 Ruili Huang 1 Misha Itkin 1 Zina Itkin 1 Paul Shinn 1 Carleen Klumpp-Thomas 1 Sam Michael 1 Carlos A Tristan 1 Anton Simeonov 1 Ilyas Singeç 2
Affiliations

Affiliations

  • 1 Stem Cell Translation Laboratory (SCTL), Division of Preclinical Innovation (DPI), National Center for Advancing Translational Sciences (NCATS), National Institutes of Health (NIH), Rockville, MD, USA.
  • 2 Stem Cell Translation Laboratory (SCTL), Division of Preclinical Innovation (DPI), National Center for Advancing Translational Sciences (NCATS), National Institutes of Health (NIH), Rockville, MD, USA. ilyas.singec@nih.gov.
Abstract

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.

Keywords

Cell viability; Dose–response curves; Embryonic stem cells; High-throughput screening; Induced pluripotent stem cells; Robotic cell culture; Small molecules; Toxicity.

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