摘要
目的观察大鼠骨髓源性间充质干细胞(MSCs)对内皮细胞(ECs)增殖及成管能力的影响。方法采用密度梯度离心、流式细胞仪及免疫荧光的方法将不同培养条件下(DMEM及EGM-2培养液)大鼠骨髓源性单核细胞(BM—MNCs)进行分离及鉴定,并对直接和间接共培养(Transwells)的细胞进行体外成管和免疫荧光统计分析。结果与DMEM中的细胞比较,EGM-2培养的BM—MNCs能够形成典型的集落形成细胞。流式细胞仪检测和免疫荧光鉴定表明培养在DMEM和EGM-2中的细胞分别为MSCs(CD105、CD90和CD73表达率大于95%)和ECs(CD31阳性率大于90%)。与间接共培养比较,直接共培养后细胞的成管数量及长度均显著降低(P〈0.05)。免疫荧光染色进一步证明,直接共培养后ECs的数量显著减少(P〈0.05)。结论直接共培养下,大鼠MSCs能够抑制ECs的增殖及成管。
Objective To investigate the effect of rat bone marrow-derived mesenchymal stem cells (MSCs) on proliferation and angiogenesis of endothelial cells (ECs). Methods The rat bone marrow-derived mononuclear cells (BM-MNCs) were collected by density gradient eentrifugation and cultured in different medium (DMEM and EGM-2) before identified by flow cytometry or immunofluorescence. After direct and indirect co-culture, they were analysised by vascular network formation in vitro and immunofluorcscence. Results Compared with DMEM, the BM-MNCs could formed typical colony-forming cells in EGM-2. After cultured in DMEM and EGM-2 respectively, MNCs could develop into MSCs with greater than 95% of cells expressing CD105, CD90 and CD73, and ECs with greater than 90% of cells positive staining for CD31. Compared with indirect co-culture, direct co-culture of MSCs and ECs cells would result in significant reduction of tube quantity and tube length (P 〈 0. 05 ). Furthermore, immunofluorescence staining demonstrated the number of ECs also significantly reduced ( P 〈 0. 05 ). Conclusion Directly co- cultured MSCs were able to inhibit ECs proliferation and vascular network formation.
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2013年第5期884-886,共3页
Chinese Journal of Experimental Surgery
关键词
大鼠骨髓干细胞
共培养
成管
增殖
Rat bone marrow stem cells
Co-culture
Vascular network formation
Proliferation