大鼠血管内皮祖细胞对骨髓间充质干细胞增殖及凋亡的影响  

Effect of rat endothelial progenitor cell on proliferation and apoptosis of bone marrow mesenchymal stem cell

作　　者：温丽[1] 温宁[2] 王宇 丁寅[4] 

机构地区：[1]解放军总医院口腔正畸科,北京100853 [2]解放军总医院口腔修复科 [3]第四军医大学第一附属医院肿瘤科 [4]第四军医大学口腔医学院正畸科,西安710032

出　　处：《中华口腔医学杂志》2014年第2期106-110,共5页Chinese Journal of Stomatology

基　　金：国家自然科学基金（81170982）

摘　　要：目的 探讨大鼠血管内皮祖细胞（endothelial progenitor cell,EPC）对骨髓间充质干细胞（bone marrow mesenchymal stem cell,BMSC）增殖及凋亡的影响,为促进BMSC再生能力提供理论依据.方法 体外分离、培养并鉴定大鼠EPC、BMSC.利用Transwell小室建立EPC、BMSC间接共培养体系,共培养组为BMSC-EPC共培养,对照组为BMSC-BMSC共培养;检测不同接种比例下（EPC：BMSC为1∶1、10∶1、100∶1）克隆集落形成率（colony forming unit-fibroblastic,CFU-F）的变化;采用流式细胞术检测间接共培养第3天,EPC对BMSC细胞周期的影响,以及共培养第3、7、10天,EPC对BMSC细胞凋亡的调控作用.用独立t检验进行两组比较,同组内不同时间点的细胞凋亡率以及不同接种比例的CFU-F比较采用单因素方差分析.结果 共培养组S期细胞比例[（15.72±2.93）％]显著高于对照组[（2.02 ±0.66）％]（P＜0.01）.不同接种比例下（10∶1、100∶1）共培养组CFU-F[（50.98±6.32）％、（57.87±14.06）％]也显著高于对照组[（33.07±9.60）％、（30.06±7.20）％,P＜0.0I].共培养第3、7、10天两组细胞凋亡率差异无统计学意义（P＞0.05）.结论 EPC可通过细胞间接接触促进BMSC增殖,但对BMSC的凋亡没有明显影响.Objective To investigate the effect of endothelial progenitor cell（EPC） on regulating proliferation and apoptosis of bone marrow mesenchymal stem cell （BMSC） in the indirect co-culture system. Methods BMSC and EPC were cultivated and identified in vitro. Using transwell inserts to establish EPC and BMSC indirect co-culture system. Experimental groups were prepared as follows： BMSC/EPC indirect co-culture： BMSC were co-cultured with EPC that were separated in transwell insert. Control： BMSC/BMSC were indirect co-cultured at the same cell counts with the experimental groups. Colony forming unit-fibroblast（CFU-F） assay was studied with three types of EPC/BMSC ratios （1： 1,10： 1,100：1 ） to assess the capacity and efficiency for cell self-renewal. In addition, flow cytometry technique was used to detect the cell cycle at co-cultured 3 days and the cell apoptosis at co-cultured 3,7,10 d. Results Under indirect co-culture condition, EPC could significantly promote cell cycle progress and enhance capacity of cell self-renewal. Co-cultured EPC resulted in an accumulation of BMSC at S phase [ experimental group： （ 15.72 ±2.93）% ,control group： （2.02 ±0. 66）% ,P 〈0. 01 ]. CFU-F assay showed that the self-renewal capacity of EPC： BMSC/10：1 group [ （ 50. 98 ± 6. 32） % 1 and 100 ： 1 group [ （ 57.87 ± 14. 06） % ] were significantly higher than that in control group [ （ 33.07 ± 9. 60 ） % and （ 30. 06 ± 7.20） % ] （ P 〈 0. 01 ）. However,EPC had a slight but non-significant down-regulated effect on BMSC apoptosis （P 〉 0. 05 ）. Conclusions Under indirect co-culture condition, EPC could enhance the proliferation of BMSC, but could not regulate cell apoptosis in vitro.

关 键 词：内皮细胞 间质干细胞 细胞增殖 细胞凋亡 共同培养技术 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]