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  3. Protocol for high-throughput viability screening and gene expression of human EndoC-βH5 β cells and pancreatic islets

Protocol for high-throughput viability screening and gene expression of human EndoC-βH5 β cells and pancreatic islets

  • STAR Protoc. 2025 Jul 18;6(3):103955. doi: 10.1016/j.xpro.2025.103955.
Nayara Rampazzo Morelli 1,  Peter J Thompson 2
Affiliations

Affiliations

  • 1 Department of Physiology and Pathophysiology, Rady Faculty of Health Sciences, University of Manitoba, Winnipeg, MB R3E 3P4, Canada; Diabetes Research Envisioned and Accomplished in Manitoba (DREAM) Theme, Children's Hospital Research Institute of Manitoba, Winnipeg, MB R3E 3P4, Canada. Electronic address: nayara.rampazzomorelli@umanitoba.ca.
  • 2 Department of Physiology and Pathophysiology, Rady Faculty of Health Sciences, University of Manitoba, Winnipeg, MB R3E 3P4, Canada; Diabetes Research Envisioned and Accomplished in Manitoba (DREAM) Theme, Children's Hospital Research Institute of Manitoba, Winnipeg, MB R3E 3P4, Canada. Electronic address: peter.thompson@umanitoba.ca.
PMID: 40682782 DOI: 10.1016/j.xpro.2025.103955
Abstract

Viability assays allow the assessment of toxicity induced by any treatment of interest. Here, we present a protocol to evaluate cell viability in human EndoC-βH5 β cells and human pancreatic islets using a high-throughput fluorescence viability assay. We describe the steps for cell and islet culture, reagent dilution, viability assay preparation, and performance. The protocol is compatible with RNA isolation and gene expression analysis on the same cells, and we detail procedures for reagent washout and profiling gene expression. For complete details on the use and execution of this protocol, please refer to Rampazzo Morelli et al.1.

Keywords

Cell Biology; Cell culture; Cell-based Assays; Metabolism.

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