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Purification and growth of endothelial progenitor cells from murine bone marrow mononuclear cells
Author(s) -
Wang Qi Ru,
Wang Bao He,
Huang Yan Hong,
Dai Guo,
Li Wei Ming,
Yan Qi
Publication year - 2007
Publication title -
journal of cellular biochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.028
H-Index - 165
eISSN - 1097-4644
pISSN - 0730-2312
DOI - 10.1002/jcb.21377
Subject(s) - bone marrow , progenitor cell , peripheral blood mononuclear cell , microbiology and biotechnology , immunology , chemistry , biology , stem cell , in vitro , biochemistry
This study reports the culture and purification of murine bone marrow endothelial progenitor cells (EPCs) using endothelial cell‐conditioned medium (EC‐CM). Endothelial‐like cells appeared at day 5 in culture of bone marrow mononuclear cells in the presence of EC‐CM in the culture system, and these cells incorporated acetylated low‐density lipoproteins (Ac‐LDL) and reacted with endothelial‐specific Ulex Europaeus Lectin . Continued incubation of these cells at low density with EC‐CM for longer than 10 days resulted in the formation of endothelial cell colonies which gave rise to colonies of endothelial progeny and can be passed for many generations in the EC‐CM culture system. Cells derived from these colonies expressed endothelial cell markers such as vWF and CD31, incorporated Dil‐Ac‐LDL, stained positive for Ulex Europaeus Lectin , formed capillary‐like structures on Matrigel, and demonstrated a high proliferative capacity in culture. These bone marrow‐derived adherent cells were identified as EPCs. The purification and the formation of EPC colonies by using EC‐CM were associated with the cytokines secreted in the EC‐CM. VEGF, bFGF, and GM‐CSF in the EC‐CM stimulated the proliferation and growth of EPCs, whereas AcSDKP (tetrapeptide NAc‐Ser‐Asp‐Lys‐Pro) in EC‐CM suppressed the growth of mesenchymal stem cells (MSC) and fibroblasts. This approach is efficient for isolation/purification and outgrowth of bone marrow EPCs in vitro, a very important cell source in angiogenic therapies and regenerative medicine. J. Cell. Biochem. 103: 21–29, 2008. © 2007 Wiley‐Liss, Inc.