骨髓间充质干细胞的贴壁培养及内毒素对其增殖活性的影响  

Adherent culturing and effects of endotoxin on proliferation of bone marrow mesenchymal stem cells

作　　者：郝彦琴[1] 卢宁[1] 李红[1] 赵龙凤[1] 

机构地区：[1]山西医科大学第一医院感染病科,太原市030001

出　　处：《中华实验和临床感染病杂志（电子版）》2010年第2期12-15,共4页Chinese Journal of Experimental and Clinical Infectious Diseases(Electronic Edition)

基　　金：山西省研究生优秀创新项目(20081047)

摘　　要：目的贴壁法分离培养大鼠骨髓间充质干细胞(BMSCs),并观察不同浓度内毒素对其增殖活性的影响。方法采用全骨髓贴壁培养法培养大鼠BMSCs,倒置显微镜下观察细胞形态,免疫细胞化学方法检测细胞CD44、CD29、CD34的表达,流式细胞术检测细胞周期。加不同浓度的内毒素(0、0.01μg/ml、0.1μg/ml、1μg/ml、10μg/ml、100μg/ml)作用24h,用四唑盐比色法(MTT)检测BMSCs的增殖。结果分离培养的细胞三代以后呈均一的成纤维细胞样形态,高表达CD44,但不表达CD34、CD45。80%以上的第三代BMSCs处于G1期。不同浓度的LPS作用24h后各组的平均吸光度值(A)比较有统计学意义(F=3.598,P=0.007);0.01μg/mlLPS组A值与其余各组相比,具有统计学意义(P<0.05)。结论全骨髓贴壁培养法可以方便、快捷地获得BMSCs,其在形态学、细胞表面标志物表达和多向分化能力方面具有干细胞生物学特性;0.01μg/mlLPS可明显促进BMSCs的增殖。Objective To isolate and culture rat bone marrow-derived mesenchymal stem cells（BMSCs）in vitro by adherent culturing and to observe the effects of different endotoxin concentrations on proliferation of rat BMSCs.Methods The BMSCs were purified and amplified by adherent culturing in vitro.The changes of cells morphology were observed with invert microscope.The cell member antigens CD34,CD44,CD45 were examined with immunocytochemical technique.Cell cycle was detected with flow cytometry.The proliferation of BMSCs was determined through MTT method after the BMSCs were treated with different LPS concentrations for 24 hours.Results The morphology of P3 BMSCs were fibroblast-like.Immunohistochemical results of BMSCs showed that CD44 was positive and CD34,CD45 were negative.About 80% of P3 BMSCs were in G1 phase.The average A values were significantly different among various LPS concentrations groups（F=3.598,P=0.007）;The average A value of 0.01 μg/ml LPS group were improved significantly than that in other groups（P0.01）.Conclusions BMSCs can be conveniently and quickly obtained by bone marrow adherent culturing.The cultured cells have general biological characteristics of stem cell in morphology and can express cell member antigens and multi-directionally differentiate.0.01 μg/ml LPS can significantly improve the proliferation of rat BMSCs.

关 键 词：内毒素 骨髓间充质干细胞 细胞增殖 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]