Glucose-Responsiveness of Pancreatic β-Like (GRP β-L) Cells Generated from Human Pluripotent Stem Cells

Curr Protoc Hum Genet. 2019 Jan;100(1):e71. doi: 10.1002/cphg.71. Epub 2018 Oct 18.

Abstract

The International Diabetic Federation estimated that 415 million adults currently have diabetes and 318 million adults had impaired glucose tolerance, putting them at high risk of developing diabetes in the future. In Type 1 Diabetes (T1D), the β cells are lost because of autoimmune reactions. Although islet transplantation has been a promising therapy for T1D, it is greatly limited by pancreatic donors. Here, we describe a protocol to generate glucose- responsive pancreatic β-like (GRPβ-L) cells from human-induced pluripotent stem (iPS) cells. We recapitulate in vivo pancreas development by in vitro induction of differentiating human (iPS) cells with stage-specific signaling molecules and proteins. Inhibition of Tyrosine Kinase receptor AXL, TGF-β, and Notch signaling pathways in the final stage of the five-stage protocol could efficiently generate GRPβ-L from the endocrine progenitor. Differentiation of human iPS cells through the protocol could result in functional GRPβ-L cells, which could be used in pharmaceutical and β cell biology studies. © 2018 by John Wiley & Sons, Inc.

Keywords: embryonic stem (ES) cell; induced pluripotent stem (iPS) cell; insulin; protocol to generate beta cells; β-like cells.

MeSH terms

  • Axl Receptor Tyrosine Kinase
  • Cell Culture Techniques / methods*
  • Cell Differentiation / genetics*
  • Cell Lineage / genetics
  • Diabetes Mellitus, Type 1 / genetics*
  • Diabetes Mellitus, Type 1 / metabolism
  • Diabetes Mellitus, Type 1 / pathology
  • Diabetes Mellitus, Type 1 / therapy
  • Glucose / metabolism
  • Humans
  • Induced Pluripotent Stem Cells / metabolism
  • Insulin-Secreting Cells / metabolism*
  • Insulin-Secreting Cells / pathology
  • Proto-Oncogene Proteins / antagonists & inhibitors
  • Receptor Protein-Tyrosine Kinases / antagonists & inhibitors
  • Receptors, Notch / antagonists & inhibitors
  • Signal Transduction
  • Transforming Growth Factor beta / antagonists & inhibitors

Substances

  • Proto-Oncogene Proteins
  • Receptors, Notch
  • Transforming Growth Factor beta
  • Receptor Protein-Tyrosine Kinases
  • Glucose
  • Axl Receptor Tyrosine Kinase
  • AXL protein, human