CAJ | 학술논문

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的数量湿著减少(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.
목적 관찰대서골수원성간충질간세포(MSCs)대내피세포(ECs)증식급성관능력적영향.방법 채용밀도제도리심、류식세포의급면역형광적방법장불동배양조건하(DMEM급EGM-2배양액)대서골수원성단핵세포(BM-MNCs)진행분리급감정,병대직접화간접공배양(Transwells)적세포진행체외성관화면역형광통계분석.결과 여DMEM중적세포비교,EGM-2배양적BM-MNCs능구형성전형적집락형성세포.류식세포의검측화면역형광감정표명배양재DMEM화EGM-2중적세포분별위MSCs(CD105、CD90화CD73표체솔대우95％)화ECs(CD31양성솔대우90％).여간접공배양비교,직접공배양후세포적성관수량급장도균현저강저(P＜0.05).면역형광염색진일보증명,직접공배양후ECs적수량습저감소(P＜0.05).결론 직접공배양하,대서MSCs능구억제ECs적증식급성관.
Objective To investigate the effect of rat bone marrow-derived mesenchvmal 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 centrifugation and cultured in different medium (DMEM and EGM-2) before identified by flow cytometry or immunofluorescenee.After direct and indirect co-culture,they were analysised by vascular network formation in vitro and immunofluorescence.Results Compared with DMEM,the BM-MNCs could formed typical colony-forming cells in EGM-2.After cuhured 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 tabe length (P ＜ 0.05).Furthermore,immunofluorescence staining demonstrated the number of ECs also significantly reduced (P ＜ 0.05).Conclusion Directly cocultured MSCs were able to inhibit ECs proliferation and vascular network formation.