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

关键词:  胎盘间充质干细胞  低氧  生物学特性

Key words:  [Key words]　human placenta mesenchymal stem cells  hypoxia  biological characteristics

摘要:

[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%O2）及不同浓度的低氧（5%O2、3%O2、1%O2、0.5%O2）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%O2~1%O2） 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%O2）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。

Abstract:

[Abstract] 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 tissue culture method and their surface markers were identified by flow cytometry. The third generation of pMSCs were cultured in the environment of normoxia (21%O2) and hypoxia of different concentrations (5%O2, 3%O2, 1%O2, 0.5% O2).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 isolated. They expressed CD29, CD44 positively but CD31，CD34，CD45，HLA-DR negatively. The morphology of pMSCs cultured in hypoxia are uniform, long spindle shapes and whorls significantly. Compared with normoxia group, the proliferation, migration and VEGF secretion of pMSCs cultured in hypoxia within a certain range (5%O2-1%O2) increased. The lower the oxygen concentration, the more obvious the changes were. However, the proliferation and VEGF secretion of pMSCs cultured in low oxygen concentration (0.5%O2) 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.