The characteristics of endothelial progenitor cells derived from mononuclear cells of rat bone marrow in different culture conditions |
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Authors: | Nana Yang Dawei Li Peng Jiao Bin Chen Shutong Yao Hui Sang Mingfeng Yang Jiju Han Ying Zhang Shucun Qin |
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Institution: | (1) Institute of Atherosclerosis, Taishan Medical University, Taian, 271000, China |
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Abstract: | Endothelial progenitor cells (EPCs) derived from bone marrow are known to be heterogeneous. In this study, we tried to find
favorable conditions that induce the differentiation of mononuclear cells (MNCs) from bone marrow into EPCs. The differentiation
capacity of MNCs from rat bone marrow was investigated in different conditions, such as different media, different induction
times and different culture surfaces. The cell morphology and endothelial biomarkers associated with differentiated MNCs were
studied. Our results indicated that MNCs cultured in EGM-2MV (Endothelial cell basal medium-2, plus SingleQuots of growth
supplements) developed a bursiform shape, a late EPC-like morphology, while MNCs cultured in complete medium (CM, M199 with
10% FBS, 20 ng/mL VEGF and 10 ng/mL bFGF) showed a spindle shape, an early EPC-like morphology. Cells of both morphologies
were able to incorporate DiI-ac-LDL and bind lectin in vitro. MNCs cultured in EGM-2MV exhibited a higher proliferation rate
and higher eNOS expression than MNCs cultured in CM. MNCs cultured in EGM-2MV had the ability to form tubes on Matrigel. Flow
cytometry results indicated that CD133 expression was highest at day 12 and that the greatest number of cells positive for
both FLK-1 and CD133 appeared at day 20 from cells cultured in dishes without fibronectin coating. In addition, the expression
levels of CD133, CD31 and FLK-1/CD133 were not significantly different between cells of different shapes. Our experiments
suggest that MNCs from bone marrow can be differentiated into late EP-like cells in EGM-2MV, which have the ability to rapidly
proliferate. These MNCs can also be differentiated into early EP-like cells in CM. Additionally, fibronectin may not be necessary
for the differentiation of EPCs to mature ECs after three generations. Differentiated MNCs from bone marrow in EGM-2MV have
the characteristics of EPCs, although the expression levels of EPC markers were lower than previously reported. |
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Keywords: | Endothelial progenitor cells Mononuclear cells CD133 FLK-1 |
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