Generation of a fluorescent mNeonGreen insulin reporter line in the H1 (WA01) hESC background

Stem Cell Res. 2024 Dec:81:103559. doi: 10.1016/j.scr.2024.103559. Epub 2024 Sep 11.

Abstract

Over the past decade, the use of human stem cell-derived β cells (SC-β cells) to model pancreatic β cell development, function and disease has become increasingly common. Though protocols are rapidly improving, current directed differentiation strategies do not yield a pure population of insulin-positive SC-β cells in vitro. Therefore, it is experimentally advantageous to have reporter lines that allow for live sorting of insulin-positive populations. To aid in these studies, we have knocked mNeonGreen fluorescent protein into the endogenous insulin locus of the commonly used H1 (WA01) human embryonic stem cell line.

MeSH terms

  • Cell Differentiation
  • Cell Line
  • Genes, Reporter*
  • Human Embryonic Stem Cells* / cytology
  • Human Embryonic Stem Cells* / metabolism
  • Humans
  • Insulin* / metabolism
  • Insulin-Secreting Cells* / cytology
  • Insulin-Secreting Cells* / metabolism
  • Luminescent Proteins / genetics
  • Luminescent Proteins / metabolism

Substances

  • Insulin
  • Luminescent Proteins