Functional β-Cell Differentiation of Small-Tail Han Sheep Pancreatic Mesenchymal Stem Cells and the Therapeutic Potential in Type 1 Diabetic Mice

Pancreas. 2020 Aug;49(7):947-954. doi: 10.1097/MPA.0000000000001604.

Abstract

Objectives: This study aims to investigate the characteristics of sheep pancreatic mesenchymal stem cells (PSCs) and therapeutic potential of differentiated β-like cells in streptozotocin-induced diabetic mice.

Methods: Pancreatic mesenchymal stem cells were isolated from 3- to 4-month-old sheep embryos, and their biological characteristics were explored. The function and therapeutic potential of differentiated β-like insulin-producing cells were also investigated in vitro and in vivo. Differentiated cells were identified through dithizone staining and immunofluorescence staining. Insulin secretion was analyzed using an enzyme-linked immunosorbent assay kit. The preliminary therapeutic potential of induced β-like cells in diabetic mice was detected by blood glucose and body weight.

Results: Primary PSCs were isolated and subcultured up to passage 36. Immunofluorescence staining presented PSC-expressed important markers such as Pdx1, Nkx6-1, Ngn3, and Nestin. Primary PSCs could be induced into functional pancreatic β-like islet cells with a 3-step protocol. The induced β-like islet cells could ameliorate blood glucose in diabetic mice.

Conclusions: The method proposed for generating pancreatic islet β cells provided a preliminary phenotypic investigation of induced cell treatment in diabetic mice, and also laid a foundation in the identification of pharmaceutical targets to treat insulin-dependent diabetes.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Blood Glucose / metabolism
  • Body Weight
  • C-Peptide / metabolism
  • Cell Differentiation*
  • Cells, Cultured
  • Diabetes Mellitus, Experimental / blood
  • Diabetes Mellitus, Experimental / therapy*
  • Diabetes Mellitus, Type 1 / blood
  • Diabetes Mellitus, Type 1 / therapy*
  • Glucagon / metabolism
  • Glucose / pharmacology
  • Humans
  • Insulin / metabolism
  • Insulin Secretion / drug effects
  • Insulin-Secreting Cells / cytology*
  • Insulin-Secreting Cells / metabolism
  • Male
  • Mesenchymal Stem Cell Transplantation / methods*
  • Mesenchymal Stem Cells / cytology*
  • Mesenchymal Stem Cells / metabolism
  • Mice
  • Sheep

Substances

  • Blood Glucose
  • C-Peptide
  • Insulin
  • Glucagon
  • Glucose