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.

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.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, N.I.H., Intramural

MeSH terms

  • Cell Culture Techniques / methods
  • Cell Differentiation / physiology
  • Drug Evaluation, Preclinical
  • High-Throughput Screening Assays / methods
  • Humans
  • Induced Pluripotent Stem Cells*
  • Pluripotent Stem Cells*