氧浓度对人胎盘间充质干细胞生物学特性的影响  

Effect of different oxygen concentration on the biological characteristics of human placenta derived mesenchymal stem cells

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作  者:杜莉莉[1] 吕润潇[2,3] 杨晓漪[4] 辛娜[1] 马廷贤[1] 

机构地区:[1]中国医科大学基础医学院病理生理学教研室,辽宁沈阳110013 [2]第二军医大学附属长海医院研究生队,上海200433 [3]第二军医大学研究生队,上海200433 [4]中国医科大学

出  处:《大连医科大学学报》2015年第5期442-446,共5页Journal of Dalian Medical University

摘  要:目的:比较不同氧浓度对人胎盘间充质干细胞生物学特性的影响。方法组织块培养法分离人胎盘间充质干细胞pMSCs,并利用流式细胞技术进行表面标志鉴定。将第3代pMSCs分别培养于常氧(21%O2)及不同浓度的低氧(5%O2、3%O2、1%O2、0.5%O2)环境中,观察各组细胞形态,MTT检测细胞增殖情况,划痕实验检测细胞迁移能力,ELISA检测细胞VEGF分泌表达量情况。结果成功从人胎盘组织分离出间充质干细胞pMSCs,阳性表达CD29、CD44,阴性表达CD31、CD34、CD45、HLA-DR。低氧培养的pMSCs形态均一,细胞伸展为长梭形,漩涡状排列明显。与常氧组相比,一定范围内低氧(5%O2~1%O2)培养的pMSCs增殖、迁移增快,分泌VEGF增多,氧浓度越低,变化越明显。过低氧浓度(0.5%O2)培养的pMSCs增殖减慢,分泌VEGF减少。结论一定范围内的低氧可促进pMSCs增殖、迁移及分泌VEGF,氧浓度越低,促进作用越明显。而过低氧浓度则产生抑制作用。Objective To compare the effects of different oxygen concentrations on the biological characteristics of human placenta derived mesenchymal stem cells.Methods Human placenta derived mesenchymal stem cells were isolated by tis-sue culture method and their surface markers were identified by flow cytometry.The third generation of pMSCs were cul-tured in the environment of normoxia (21%O2 ) and hypoxia of different concentrations (5%O2 , 3%O2 , 1%O2 , 0.5%O2 ) .The morphologic changes of pMSCs were detected by inverted phase contrast microscope.The proliferation of pMSCs were examined by methyl-tetrazolium ( MTT) assay.The migration of pMSCs were analyzed by scratch assay and the VEGF secretion was detected by ELISA.Results Human placenta derived mesenchymal stem cells were successfully isola-ted.They expressed CD29, CD44 positively but CD31,CD34,CD45,HLA-DR negatively.The morphology of pMSCs cul-tured in hypoxia are uniform, long spindle shapes and whorls significantly.Compared with normoxia group, the prolifera-tion, migration and VEGF secretion of pMSCs cultured in hypoxia within a certain range (5%O2 -1%O2 ) increased.The lower the oxygen concentration, the more obvious the changes were.However, the proliferation and VEGF secretion of pM-SCs cultured in low oxygen concentration (0.5%O2 ) decreased.Conclusions Hypoxia within a certain range can promote proliferation, migration and VEGF secretion of pMSCs.The lower the oxygen concentration, the more obvious the promoting effect is.But too low oxygen concentration produces inhibition effect.

关 键 词:胎盘间充质干细胞 低氧 生物学特性 

分 类 号:R329.2[医药卫生—人体解剖和组织胚胎学]

 

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