两种不同内皮祖细胞的分离培养与鉴定

发布时间：2018-07-23 20:15

【摘要】：目的:探讨从小鼠骨髓中分离、培养、诱导分化及鉴定两种内皮祖细胞的方法,为进一步研究和临床应用奠定基础。方法:密度梯度离心法分离小鼠骨髓单个核细胞,接种于内皮祖细胞条件培养基,通过贴壁培养法培养出早期内皮祖细胞和晚期内皮祖细胞,并在0 d、6 d、10 d流式鉴定早期内皮祖细胞,在第8周流式鉴定晚期内皮祖细胞。结果:通过体外贴壁扩增培养,从小鼠骨髓细胞中成功培养出EEPC(早期内皮祖细胞)和EOC(晚期内皮祖细胞),表达CD34+/CD133+/VEGFR2+的EEPC比例从最初的0.08%能够增长至70%;EOC大约出现于3-4周,5-8周时呈现指数增长,具有典型的内皮细胞鹅卵石样形态,表达CD31、VEGFR2等内皮细胞表面标志而不表达CD34、CD133等干细胞表面标志。结论:确立了内皮祖细胞体外分离培养和诱导分化的实验方法,为进一步研究奠定基础。
[Abstract]:Objective: to investigate the methods of isolation, culture, differentiation and identification of two kinds of endothelial progenitor cells from mouse bone marrow, so as to lay a foundation for further study and clinical application. Methods: murine bone marrow mononuclear cells were isolated by density gradient centrifugation and inoculated in the conditioned medium of endothelial progenitor cells. The early and late endothelial progenitor cells were cultured by adherent culture. Endothelial progenitor cells (EPCs) were identified by flow cytometry at 0 d, 6 d, 10 d, and 8 weeks, respectively. Results: through in vitro adherent amplification culture, EEPC (early endothelial progenitor cells) and EOC (late endothelial progenitor cells) were successfully cultured from mouse bone marrow cells. The proportion of EEPC expressing CD34 / CD133 / VEGFR2 increased from 0.08% to 70%. It expressed CD31VEGFR2 and other endothelial cell surface markers, but not CD34, CD133 and other stem cell surface markers. Conclusion: the method of isolation, culture and differentiation of endothelial progenitor cells in vitro was established, which laid a foundation for further study.
【作者单位】：第二炮兵工程大学门诊部;第四军医大学第二附属医院唐都医院血液内科;
【分类号】：R329.2;Q813

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