中华医学杂志
中華醫學雜誌
중화의학잡지
National Medical Journal of China
2010年
30期
2149-2152
,共4页
宋玮玮%白海%王存邦%鱼丽莉%欧剑锋%赵强%苏亚楠
宋瑋瑋%白海%王存邦%魚麗莉%歐劍鋒%趙彊%囌亞楠
송위위%백해%왕존방%어려리%구검봉%조강%소아남
间质干细胞%细胞低氧%细胞增殖%人
間質榦細胞%細胞低氧%細胞增殖%人
간질간세포%세포저양%세포증식%인
Mesenchymal stem cells%Cell hypoxia%Cell proliferation%Persons
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.
目的 探討二氯化鈷(CoCl2)誘導的化學性低氧對人骨髓間充質榦細胞(hBMSC)增殖的影響.方法 採用密度梯度離心法分離、培養hBMSC;用流式細胞儀檢測其錶麵標誌物;用CoCl2建立化學性缺氧模型,分彆採用四甲基偶氮唑鹽(MTT)法和流式細胞儀檢測hBMSC在不同CoCl2濃度(0、100、150、300 μmol/L)和不同時間(0,12 h及1、2、4、6、7 d)的增殖情況,0 μmol/L CoCl2組為常氧組,後3箇濃度組為低氧組.結果 化學缺氧12 h內,hBMSC增殖被抑製;1、2、4 d時各低氧組細胞增殖均高于常氧組,但300 μmol/L CoCl2組與常氧組比差異無統計學意義(P＞0.05);而100μmol/L CoCl2組(0.139±0.003、0.178±0.005、0.224±0.005)和150 μmol/L CoCl2組(0.202±0.020、0.224±0.019、0.263±0.004)增殖明顯高于常氧組(0.134±0.005、0.167±0.004、0.206±0.005),其中150μmol/L CoCl2組1 d時增殖最顯著(均P＜0.05).6 d時100、150 μmol/LCoCl2組細胞增殖(0.258±0.020、0.264±0.008)仍高于常氧組(0.248±0.004),但優勢逐漸減弱(P＜0.05).7 d時這種低氧促增殖的效應消失.結論 CoCl2誘導的化學性缺氧可以促進hBMSC增殖.
목적 탐토이록화고(CoCl2)유도적화학성저양대인골수간충질간세포(hBMSC)증식적영향.방법 채용밀도제도리심법분리、배양hBMSC;용류식세포의검측기표면표지물;용CoCl2건립화학성결양모형,분별채용사갑기우담서염(MTT)법화류식세포의검측hBMSC재불동CoCl2농도(0、100、150、300 μmol/L)화불동시간(0,12 h급1、2、4、6、7 d)적증식정황,0 μmol/L CoCl2조위상양조,후3개농도조위저양조.결과 화학결양12 h내,hBMSC증식피억제;1、2、4 d시각저양조세포증식균고우상양조,단300 μmol/L CoCl2조여상양조비차이무통계학의의(P＞0.05);이100μmol/L CoCl2조(0.139±0.003、0.178±0.005、0.224±0.005)화150 μmol/L CoCl2조(0.202±0.020、0.224±0.019、0.263±0.004)증식명현고우상양조(0.134±0.005、0.167±0.004、0.206±0.005),기중150μmol/L CoCl2조1 d시증식최현저(균P＜0.05).6 d시100、150 μmol/LCoCl2조세포증식(0.258±0.020、0.264±0.008)잉고우상양조(0.248±0.004),단우세축점감약(P＜0.05).7 d시저충저양촉증식적효응소실.결론 CoCl2유도적화학성결양가이촉진hBMSC증식.
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.
