A simple method of hiPSCs differentiation into insulin-producing cells is improved with vitamin C and RepSox

PLoS One. 2021 Jul 12;16(7):e0254373. doi: 10.1371/journal.pone.0254373. eCollection 2021.

Abstract

Human induced pluripotent stem cells (hiPSCs) are considered a promising source of pancreatic β-cells for the treatment of diabetes. However, this approach is limited by issues such as low efficiency and high cost. Here, we have developed a new protocol to induce insulin-producing cells. To reduce costs, we decreased the number of reagents and replaced protein reagents with chemical compounds. In this method, we increased induction efficiency with ascorbic acid (vitamin C) and an ALK5 inhibitor, RepSox. In 2D culture, the majority of cells were immature β-cells with low glucose-stimulated insulin secretion. Transferring to 3D culture immediately after endocrine progenitor cell differentiation, however, improved glucose-stimulated insulin secretion. This simplified method will contribute to realizing transplantation therapy of β-cells using iPSCs.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Ascorbic Acid / pharmacology*
  • Cell Count
  • Cell Culture Techniques / methods*
  • Cell Differentiation*
  • Endoderm / cytology
  • Glucose / pharmacology
  • Humans
  • Induced Pluripotent Stem Cells / cytology*
  • Insulin Secretion / drug effects
  • Insulin-Secreting Cells / cytology*
  • Mice
  • Pyrazoles / pharmacology*
  • Pyridines / pharmacology*

Substances

  • Pyrazoles
  • Pyridines
  • RepSox
  • Glucose
  • Ascorbic Acid

Grants and funding

Research Center Network for Realization of Regenerative Medicine program of Japan agency for Medical Research and Development (16bm0304005h0004 to T.M.), MEXT/JSPS KAKENHI (19K16138 to T.Y.).