Biological properties of bone marrow-derived early and late endothelial progenitor cells in different culture media
Biological properties of bone marrow-derived early and late endothelial progenitor cells in different culture media
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不同培养基中骨髓来源的早期和晚期内皮祖细胞的生物学特性。
DOI：
10.3892/mmr.2013.1718
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发表时间：
2013-12-01
影响因子：
3.4
通讯作者：
Zhang, Xiao Y.
中科院分区：
文献类型：
--
作者：
Guan, Xiu M.;Cheng, Min;Zhang, Xiao Y.
内皮祖细胞的体外扩增可能是克服细胞数量有限的临床问题的一种有前途的策略。由于培养基是决定细胞特性的关键，本研究探讨了不同培养基对内皮祖细胞的影响。大鼠骨髓单个核细胞在不同的培养液中培养，包括M-199含20%胎牛血清(FBS)和牛垂体提取物(M1)、M-199含10%胎牛血清(FBS)、20 ng/ml血管内皮生长因子(VEGF)和10 ng/ml碱性成纤维细胞生长因子(bFGF；M2)或表皮生长因子(EGM)-2 MV培养液。然后在体外检测细胞形态和生物学功能，如增殖、黏附、迁移、管状形成和一氧化氮(NO)的产生。此外，还对内皮生物标志物和细胞凋亡进行了分析。结果表明，所有培养体系中均可见内皮样细胞。第一代细胞即早期内皮祖细胞在M2和EGM-2 MV培养液中倾向于形成集落，而在M1培养液中呈纺锤形。在EGM-2 MV培养液中培养的第三代或第四代内皮祖细胞，即晚期内皮祖细胞，与在M1或M2培养液中培养的内皮祖细胞相比，表现出更多的黏附、迁移、管状形成和不产生。此外，在EGM-2 MV中培养的晚期内皮祖细胞表达较高水平的内皮细胞标志物，如von威廉因子(VWF)和CD31，但观察到相对较高的凋亡水平。总之，细胞培养条件，例如所使用的培养液，会影响早期和晚期骨髓来源的内皮祖细胞的生物学特性。
Ex vivo expansion of endothelial progenitor cells (EPCs) may be a promising strategy to overcome the clinical problem of limited cell numbers. As the culture medium is the key for the cell characteristics, the effects of different culture media on EPCs were investigated in the present study. Rat bone marrow mononuclear cells were cultured in different media, including M-199 media with 20% fetal bovine serum (FBS) and bovine pituitary extract (M1); M-199 media with 10% FBS, 20 ng/ml vascular endothelial growth factor (VEGF) and 10 ng/ml basic fibroblast growth factor (bFGF; M2) or epidermal growth medium (EGM)-2MV media. The cell morphology and biological functions, such as proliferation, adhesion, migration, tube formation and nitric oxide (NO) production were subsequently assayed in vitro. Moreover, endothelial biomarkers and apoptosis were also analyzed. The results showed that endothelial‑like cells appeared in all of the culture systems. First‑passage cells, namely early EPCs, tended to form colonies in M2 and EGM-2MV media but showed a fusiform shape in M1 media. The 3rd or 4th generation EPCs, namely late EPCs, cultured in EGM-2MV media exhibited increased adhesion, migration, tube formation and NO production as compared with EPCs in M1 or M2 media. Furthermore, late EPCs cultured in EGM-2MV expressed higher levels of endothelial cell markers, such as von Willibrand factor (vWF)and CD31, but relatively greater levels of apoptosis were observed. In conclusion, cell culture conditions, for example the medium used, affects the biological properties of bone marrow-derived early and late EPCs.