Hypoxia contributes to directed differentiation of mice bone marrow derived mesenchymal stem cells

doi:10.3969/j.issn.1006-7795.2015.01.004

Abstract

Abstract: Objective To monitor the intravital oxygen tension in mice femoral marrow, and assess effect of different oxygen tension on bone marrow derived mesenchymal stem cells (BM-MSCs). Methods Oxygen tension of mice femoral bone marrow was monitored by micro electrode to determine hypoxic level for ex-vivo cell culture. Mice BM-MSCs were obtained by density gradient centrifugation and cultured under 3% oxygen (hypoxia) and 20% (normoxia) oxygen in endothelial growth media-2(EGM-2). Dil-Ac-LDL and FITC-UEA absorbance assays were applied on 8 days culture of Passage 2. MTT proliferation and adhesion assays were performed on those endothelial-like cells under different oxygen condition. Results The oxygen tension in normal femoral bone marrow was (21.55±3.4)mmHg (1 mmHg=0.133 kPa), which equaled to (2.8±0.45)% (approximately 3%) oxygen in volume fraction. Mice BM-MSCs differentiated to endothelial-like cells in both 3% (hypoxia) and 20% (normoxia) oxygen. OA values for MTT proliferation assay was 0.31±0.05 and 0.16±0.01 in hypoxia and normoxia, respectively (P<0.01, n=5) and adhesion rate of endothelial-like cells was (8.72±2.95)% and (3.91±0.72)%, respectively (P<0.05, n=5). Conclusion Physiologic oxygen concentration in marrow is approximately 3%. At this level, directed differentiated endothelial-like cells present more proliferative and adhesive abilities.

Key words: oxygen partial pressure, mesenchymal stromal cells, hypoxia

CLC Number:

R459.6

Huang Ying, Zhang Jian, Chen Bing, Guo Lianrui, Li Jianxin, Zhang Shuwen. Hypoxia contributes to directed differentiation of mice bone marrow derived mesenchymal stem cells[J]. Journal of Capital Medical University, 2015, 36(1): 18-22.

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URL: https://journal03.magtech.org.cn/Jweb_sdykdxxb/EN/10.3969/j.issn.1006-7795.2015.01.004

https://journal03.magtech.org.cn/Jweb_sdykdxxb/EN/Y2015/V36/I1/18

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