Open AccessExpansion of Endothelial Progenitor Cells in High Density Dot Culture of Rat Bone Marrow Cells
Author(s) -
Lu Yang,
Yiyi Gong,
Jie Lian,
Ling Wang,
James D. Kretlow,
Guangdong Zhou,
Yilin Cao,
Wei Liu,
Wen Jie Zhang
Publication year - 2014
Publication title -
plos one
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.99
H-Index - 332
ISSN - 1932-6203
DOI - 10.1371/journal.pone.0107127
Subject(s) - bone marrow , progenitor cell , angiogenesis , microbiology and biotechnology , stem cell , endothelial stem cell , cell culture , paracrine signalling , endothelial progenitor cell , biology , in vitro , chemistry , immunology , cancer research , biochemistry , genetics , receptor
In vitro expansion of endothelial progenitor cells (EPCs) remains a challenge in stem cell research and its application. We hypothesize that high density culture is able to expand EPCs from bone marrow by mimicking cell-cell interactions of the bone marrow niche. To test the hypothesis, rat bone marrow cells were either cultured in high density (2×10 5 cells/cm 2 ) by seeding total 9×10 5 cells into six high density dots or cultured in regular density (1.6×10 4 cells/cm 2 ) with the same total number of cells. Flow cytometric analyses of the cells cultured for 15 days showed that high density cells exhibited smaller cell size and higher levels of marker expression related to EPCs when compared to regular density cultured cells. Functionally, these cells exhibited strong angiogenic potentials with better tubal formation in vitro and potent rescue of mouse ischemic limbs in vivo with their integration into neo-capillary structure. Global gene chip and ELISA analyses revealed up-regulated gene expression of adhesion molecules and enhanced protein release of pro-angiogenic growth factors in high density cultured cells. In summary, high density cell culture promotes expansion of bone marrow contained EPCs that are able to enhance tissue angiogenesis via paracrine growth factors and direct differentiation into endothelial cells.