人胎盘来源间充质干细胞和大鼠骨髓间充质干细胞的生物学特征比较

Comparisons of biological characteristics of human placenta-derived mesenchymal stem cells and rat bone marrow mesenchymal stem cells

目的:从人足月的胎盘羊膜中和Wistar大鼠股骨及胫骨骨髓中分离、培养间充质干细胞(MSCs),研究人胎盘来源MSCs(HPMSCs)和大鼠骨髓来源MSCs(BMSCs)的分离培养方法和生物学特征、表面标志及其多分化潜能。方法:将人足月胎盘组织通过胶原酶Ⅱ消化培养获取HPMSCs,采用全骨髓贴壁法从4周龄Wistar大鼠双侧股骨及胫骨中分离、纯化大鼠BMSCs,运用活细胞计数法检测其增殖能力,采用流式细胞术检测2种细胞表面标志物的表达阳性率;用地塞米松、维生素C和β磷酸甘油诱导其向成骨细胞分化,用茜素红染色鉴定;用胰岛素、地塞米松、IBMX和吲哚美辛诱导其向脂肪细胞分化,以油红O染色鉴定。结果:HPMSCs为梭形贴壁细胞,增殖能力较强,CD44和CD34表达阳性率分别为94.45%和1.67%;BMSCs为圆形、梭形和多角形,增殖能力强,CD44和CD34表达阳性率分别为93.11%和2.68%。2种细胞经过成脂诱导液和成骨诱导液诱导后,茜素红和油红O染色结果均为阳性,表明2种细胞均可向脂肪细胞和成骨细胞分化。结论:HPMSCs与大鼠BMSCs的生物学特征相似,同样具有多分化潜能,HPMSCs具有更强的增殖能力。

Objective To isolate mesenchymal stem cells（MSCs） from the amniotic membrane of human term placenta or the femur and tibia of Wistar rats and to study the methods of isolation and cultivation,biological characteristics,surface markers and multidirectional differentiation potential of human placenta-derived MSCs（HPMSCs） and rat bone marrow mesenchymal stem cells（BMSCs）.Methods The HPMSCs were isolated from human term placenta tissue by digestion of collagenase Ⅱ.The BMSCs from the femur and tibia of Wistar rats aged 4 weeks were isolated,cultured and purified by the whole bone marrow adherence method.The proliferation ability was detected by living cell number counting method.The positive expression rates of the surface markers of two kinds of cells were detected by flow cytometry.The cells were induced into osteoblasts with dexamethasone,vintamin C,and β sodium glycerophosphate;and they were also induced into adipocytes with insulin,dexamethasone,IBMX,and indometacin.After induction,the cells were observed by alizarin red staining and oil red O staining.Results HPMSCs were spindle-shaped adherent cells with strong proliferation ability.The positive expression rates of CD44 and CD34 were 94.45% and 1.67%,respectively.The MSCs isolated from the femur and tibia of rats were rounded,spindle-shaped and polygonal adherent cells with strong proliferation ability.The positive expression rates of CD44 and CD34 were 93.10% and 2.68%,respectively.After induction,the results of alizarin red staining and oil red O staining were both positive.Conclusion HPMSCs have similar biological characteristics to rat BMSCs.They also have multidirectional differentiation potential,but HPMSCs have stronger proliferation ability.