Isolation and characterization of human islet stellate cells

Exp Cell Res. 2016 Feb 1;341(1):61-66. doi: 10.1016/j.yexcr.2015.11.002. Epub 2015 Nov 4.

Abstract

Background and aims: We have previously demonstrated that islet stellate cells (ISCs) exhibiting a similar phenotype to classical pancreatic stellate cells (PSCs) could be isolated from rat islets, where they may contribute to islet fibrosis in type 2 diabetes mellitus (T2DM). This study was designed to determine whether human islets also contain ISC.

Materials and methods: Using standard explants techniques, human ISCs were enriched from freshly isolated human islets. Immunofluorescence visualization of markers for PSCs(α-smooth muscle actin;α-SMA), desmin, vimentin, glial fibrillary acidic protein (GFAP) was used to characterize the human ISC. Cell counting kit-8 (CCK-8) was used to assess the proliferation of ISC. The wound-healing assay and the transwell migration were used to assess the migration capacity of ISC. Immunofluorescence against collagen typesI (col-I), collagen typesIII (col-III) and fibronectin (FN) was performed to identify extracellular matrix (ECM) component synthesized by ISC. Adipogenic and osteogenic differentiation were tried to detected stem cell potential.

Results: In culture, ISC with triangular shape grow out from human islets. The passaged ISC expressed α-SMA, desmin, vimentin, GFAP and was positive for col-I, col-III and FN. The proliferation and migration ability of ISC was significantly slower than those of PSC. And both the human PSC and ISC were able to differentiate in vitro into adipocyte- and osteoblast-like cells.

Conclusion: Similar to our previous rat experiment, the current study shows that human islets also contain ISC which is phenotypically similar but not identical to human PSC.

Keywords: Human islet; Islet stellate cell; Type 2 diabetes mellitus.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adipogenesis
  • Biomarkers / analysis
  • Cell Culture Techniques
  • Cell Differentiation
  • Cell Movement
  • Cell Proliferation
  • Cell Separation*
  • Humans
  • Islets of Langerhans / cytology*
  • Osteogenesis
  • Pancreatic Stellate Cells / cytology*
  • Pancreatic Stellate Cells / metabolism*

Substances

  • Biomarkers