低氧对人骨髓间充质干细胞增殖的影响

目的 探讨二氯化钴(CoCl2)诱导的化学性低氧对人骨髓间充质干细胞(hBMSC)增殖的影响.方法 采用密度梯度离心法分离、培养hBMSC;用流式细胞仪检测其表面标志物;用CoCl2建立化学性缺氧模型,分别采用四甲基偶氮唑盐(MTT)法和流式细胞仪检测hBMSC在不同CoCl2浓度(0、100、150、300 μmol/L)和不同时间(0,12 h及1、2、4、6、7 d)的增殖情况,0 μmol/L CoCl2组为常氧组,后3个浓度组为低氧组.结果 化学缺氧12 h内,hBMSC增殖被抑制;1、2、4 d时各低氧组细胞增殖均高于常氧组,但300 μmol/L CoCl2组与常氧组比差异无统计学意义(P＞0.05);而100μmol/L CoCl2组(0.139±0.003、0.178±0.005、0.224±0.005)和150 μmol/L CoCl2组(0.202±0.020、0.224±0.019、0.263±0.004)增殖明显高于常氧组(0.134±0.005、0.167±0.004、0.206±0.005),其中150μmol/L CoCl2组1 d时增殖最显著(均P＜0.05).6 d时100、150 μmol/LCoCl2组细胞增殖(0.258±0.020、0.264±0.008)仍高于常氧组(0.248±0.004),但优势逐渐减弱(P＜0.05).7 d时这种低氧促增殖的效应消失.结论 CoCl2诱导的化学性缺氧可以促进hBMSC增殖.

Effects of hypoxia on the proliferation of human bone marrow mesenchymal stem cells

Objective To study the effect of hypoxia on the proliferation of human bone marrow mesenchymal stem cells (hBMSCs). Methods The method of density gradient centrifugation was employed to isolate and culture hBMSCs. And flow cytometry ( FCM) was employed to detect the cell surface marker. After establishing the experimental model of CoCl2-chemical hypoxia, MTT method and flow cytometry were applied to evaluate the proliferation and the proliferation index of hBMSCs at different time points with various CoCl2 concentrations. Results The proliferations of hBMSCs was inhibited within the first 12 hours under chemical hypoxia condition. Compared with the normal group, the hBMSCs of each CoCl2 group were remarkably proliferated 1 ,2,4 days after chemical hypoxia with CoCl2,but 300 u,mol/L CoCl2 group showed no significant difference (P >0.05). 100 μmol/L CoCl2 group (0. 139 ±0. 003, 0. 178 ±0. 005, 0. 224 ±0. 005) and 150 μmol/L CoCl2 group (0. 202 ± 0.020, 0. 224 + 0.019, 0. 263 ± 0. 004) proliferation was significantly higher than that of the control group (0. 134 ± 0. 005, 0. 167 ± 0. 004, 0. 206 ± 0. 005 ) . Compared with the normal group, the hBMSCs were remarkably proliferated 24 hours after chemical hypoxia with CoCl2 concentration of 150 )μmol/L (all P < 0.05). At Day 6 , the 100,150 u.mol/1. CoCl2 group (0. 258 ±0. 020, 0. 264 ±0.008) cells was still higher than that of normal group (0. 248 ±0. 004), but the advantage gradually diminished ( P < 0.05 ) .At Day 7 , the proliferative effects of hypoxia disappeared. Conclusion CoCl2 -induced chemical hypoxia may promote the proliferation of hBMSCs.