Isolation of mouse pancreatic ductal progenitor cells expressing CD133 and c-Met by flow cytometric cell sorting

Gastroenterology. 2007 Feb;132(2):720-32. doi: 10.1053/j.gastro.2006.11.027. Epub 2006 Nov 18.

Abstract

Background & aims: Islet transplantation has become available across the globe since a novel protocol was reported. However, because donors are in short supply, only a minority of patients benefit from this procedure. Pancreatic progenitor cells are a promising resource for regeneration of new islets, but whether progenitor cells reside in ductal epithelium is not clear.

Methods: Mouse pancreas was examined by immunohistochemistry with cell surface markers specific for ductal cells. We developed an isolation method for ductal cells by flow cytometric cell sorting using a newly identified specific marker for ductal cells. By using an in vitro colony assay, we characterized their proliferative and multipotent capacity.

Results: CD133 is expressed specifically in ductal epithelium. Flow cytometric analysis revealed that purified ductal cells are highly enriched in the CD133(+)CD34(-)CD45(-)Ter119(-) fraction. An analysis of clonal epithelial colonies formed by individual cells revealed that progenitor cells with multilineage differentiation capacity are present in neonatal ductal epithelium. Moreover, these progenitor cells express c-Met. In adult mice, progenitor cells that show a high proliferative capacity but appear committed to a ductal lineage are co-purified with CD133(+)CD34(-)CD45(-)Ter119(-) cells.

Conclusions: We established a system for isolating and culturing mouse pancreatic ductal cells that relies on flow cytometric cell sorting. Clonal analysis revealed that a population of progenitor cells is present among CD133(+) ductal cells. Isolation of these cells will facilitate future studies into the roles of pancreatic progenitor cells in regeneration and carcinogenesis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • AC133 Antigen
  • Age Factors
  • Animals
  • Animals, Newborn
  • Antigens, CD / analysis*
  • Cell Differentiation
  • Cell Lineage
  • Cell Proliferation
  • Cell Separation / methods*
  • Cells, Cultured
  • Colony-Forming Units Assay
  • Epithelial Cells / chemistry
  • Epithelial Cells / cytology*
  • Epithelial Cells / immunology
  • Flow Cytometry / methods*
  • Glycoproteins / analysis*
  • Mice
  • Mice, Inbred C57BL
  • Pancreas / chemistry
  • Pancreas / embryology
  • Pancreas / immunology
  • Pancreatic Ducts / chemistry
  • Pancreatic Ducts / cytology*
  • Pancreatic Ducts / immunology
  • Peptides / analysis*
  • Proto-Oncogene Proteins c-met / analysis*
  • Stem Cells / chemistry
  • Stem Cells / cytology*
  • Stem Cells / immunology
  • Time Factors

Substances

  • AC133 Antigen
  • Antigens, CD
  • Glycoproteins
  • Peptides
  • Prom1 protein, mouse
  • Proto-Oncogene Proteins c-met