Ex vivo generation of a highly potent population of circulating angiogenic cells using a collagen matrix

J Mol Cell Cardiol. 2011 Aug;51(2):187-97. doi: 10.1016/j.yjmcc.2011.04.011. Epub 2011 May 4.

Abstract

Biomaterials that have the ability to augment angiogenesis are highly sought-after for applications in regenerative medicine, particularly for revascularization of ischemic and infarcted tissue. We evaluated the culture of human circulating angiogenic cells (CAC) on collagen type I-based matrices, and compared this to traditional selective-adhesion cultures on fibronectin. Culture on a collagen matrix supported the proliferation of CD133(+) and CD34(+)CD133(+) CACs. When subjected to serum starvation, the matrix conferred a resistance to cell death for CD34(+) and CD133(+) progenitors and increased phosphorylation of Akt. After 4days of culture, phenotypically enriched populations of endothelial cells (CD31(+)CD144(+)) and progenitor cells (CD34(+)CD133(+)) emerged. Culture on matrix upregulated the phosphorylation and activation of ERK1/2 pathway members, and matrix-cultured cells also had an enhanced functional capacity for adhesion and invasion. These functional improvements were abrogated when cultured in the presence of ERK inhibitors. The formation of vessel-like structures in an angiogenesis assay was augmented with matrix-cultured cells, which were also more likely to physically associate with such structures compared to CACs taken from culture on fibronectin. In vivo, treatment with matrix-cultured cells increased the size and density of arterioles, and was superior at restoring perfusion in a mouse model of hindlimb ischemia, compared to fibronectin-cultured cell treatment. This work suggests that a collagen-based matrix, as a novel substrate for CAC culture, possesses the ability to enrich endothelial and angiogenic populations and lead to clinically relevant functional enhancements.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Arterioles / cytology
  • Arterioles / drug effects
  • Arterioles / metabolism
  • Biocompatible Materials / metabolism
  • Biocompatible Materials / pharmacology
  • Cell Adhesion / physiology
  • Cell Culture Techniques
  • Cell Proliferation / drug effects
  • Cell Survival / drug effects
  • Collagen / metabolism*
  • Collagen / pharmacology
  • Cytokines / blood
  • Extracellular Matrix / metabolism*
  • Extracellular Signal-Regulated MAP Kinases / metabolism
  • Gene Expression Regulation / drug effects
  • Hindlimb / blood supply
  • Humans
  • Ischemia / metabolism
  • Ischemia / therapy
  • Mesenchymal Stem Cell Transplantation
  • Mesenchymal Stem Cells / drug effects
  • Mesenchymal Stem Cells / metabolism*
  • Mice
  • Mice, Nude
  • Neovascularization, Physiologic / physiology*
  • Reperfusion

Substances

  • Biocompatible Materials
  • Cytokines
  • Collagen
  • Extracellular Signal-Regulated MAP Kinases