Isolation and large scale expansion of adult human endothelial colony forming progenitor cells

J Vis Exp. 2009 Oct 28:(32):1524. doi: 10.3791/1524.

Abstract

This paper introduces a novel recovery strategy for endothelial colony forming progenitor cells (ECFCs) from heparinized but otherwise unmanipulated adult human peripheral blood within a mean of 12 days. After large scale expansion >1x10(8) ECFCs can be obtained for further tests. Advantageously by using pHPL the contact of human cells with bovine serum antigens can be excluded. By flow cytometry and immunohistochemistry the isolated cells can be characterized as ECFC and their in vitro functionality to form vascular like structures can be tested in a matrigel assay. Further these cells can be subcutaneously injected in a mouse model to form functional, perfused vessels in vivo. After long term expansion and cryopreservation proliferation, function and genomic stability appear to be preserved. (3,4) This animal-protein free isolation and expansion method is easily applicable to generate a large quantity of ECFCs.

Publication types

  • Video-Audio Media

MeSH terms

  • Adult
  • Animals
  • Blood Cells / cytology*
  • Cell Separation / methods
  • Colony-Forming Units Assay
  • Endothelial Cells / cytology*
  • Flow Cytometry
  • Humans
  • Immunohistochemistry
  • Mice
  • Stem Cells / cytology*