Abstract
BackgroundEndothelial progenitor cells (EPCs) are increasingly becoming a major focus of regenerative medicine research and practice. The present study was undertaken to establish an appropriate procedure for isolation and characterization of EPCs from Rhesus monkeys for regenerative medicine research.ResultSelective CD34+ and nonselective mononuclear EPCs were isolated from bone marrow and cultured under varying conditions. The results showed that nonselective mononuclear EPCs were a better choice for high yield of the target cells. The cells grew in M 200 better than in EGM-2, and supplementation with fetal bovine serum promoted cell proliferation; but serum level at 7.5% was better than at 10%. In addition, surface coating of the culture dishes with human fibronectin significantly improved the proliferation and ontogeny of the isolated EPCs. Immunocytochemistry including detection of markers CD34, CD133 and CD31 and double-staining for Ac-LDL and lectin verified the purity of the cultured mononuclear EPCs.ConclusionBy a thorough analysis, we established a practical procedure for isolation and propagation of EPCs from Rhesus monkeys. This procedure would help using these valuable cells for regenerative medicine research.
Highlights
Endothelial progenitor cells (EPCs) are increasingly becoming a major focus of regenerative medicine research and practice
It is unknown for the precise ontogeny and lineage of these cells, the true extent to which EPCs participate in neovascularization and vascular repair, and the efficacy of EPC-based regenerative therapies [11,12,13]
Effects of isolation procedures on the yield and characterization of EPCs isolated from Rhesus monkeys Mononuclear EPCs isolated from bone marrow by gradient centrifugation were either directly cultured or subjected to CD34-affinity column for further purification
Summary
Endothelial progenitor cells (EPCs) are increasingly becoming a major focus of regenerative medicine research and practice These cells are enriched in the mononuclear cell fraction of peripheral blood but have been isolated from bone marrow, the vessel wall, and a number of other organs and tissues [1,2,3]. Both experimental [4,5,6,7] and human clinical trials of EPC-based therapies [8,9,10] have generated encouraging results that underscored the significance of this cell type in cardiovascular medicine; a role for EPCs in the modulation of angiogenesis has been recognized [1,2,3]. It is unknown for the precise ontogeny and lineage of these cells, the true extent to which EPCs participate in neovascularization and vascular repair, and the efficacy of EPC-based regenerative therapies [11,12,13].
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