目的:探讨氯化钴(CoCl2)模拟缺氧预处理对人脐带间充质干细胞(UC-MSCs)中碱性成纤维生长因子(bFGF)和血管内皮生长因子(VEGF)基因表达及蛋白分泌的影响。方法:分离、培养UC-MSCs。取第3代的UC-MSCs,运用含150μmol/L的CoCl2培养液培养细胞72 h,用流式细胞仪检测缺氧对UC-MSCs表面标志物(CD34、CD45、CD90、CD105、CD29及CD49)的影响。依据CoCl2的浓度(0、50、100μmol/L及150μmol/L)将UC-MSCs分为4个组,即对照组、低、中、高CoCl2组,每个组处理24 h、48 h和72 h。用SYBRGREN荧光定量RT-PCR检测bFGF、VEGF mRNA的表达,用ELISA法检测bFGF和VEGF蛋白的表达。结果:经含150μmol/L CoCl2的培养液缺氧处理,没有改变细胞的形态和表面标记物。在缺氧培养时间相同的情况下,bFGF、VEGF基因的表达均随着CoCl2浓度的增加而增加,CoCl2为150μmol/L时达到峰值(P<0.05)。当CoCl2浓度为150μmol/L培养不同时间(24 h、48 h及72 h)时,随着缺氧预处理时间的延长,bFGF、VEGF基因的表达随之增加,于48 h时达到峰值(P<0.05),bFGF、VEGF基因的表达分别为8.15±0.10和16.67±0.17,延长缺氧培养的时间基因的表达减小(P<0.05);蛋白与基因表达的变化基本一致。在含150μmol/L CoCl2的培养液培养48 h,bFGF和VEGF的表达达到顶峰(P<0.05),分别为(69.63±7.90)ng/L和(89.55±5.45)ng/L。结论:适当浓度的CoCl2(<150μmol/L)对UC-MSCs的生物学功能影响轻微。CoCl2可用于细胞的模拟缺氧,缺氧对UC-MSCs bFGF、VEGF的分泌具有时间和剂量依赖性,以含150μmol/L CoCl2的培养液培养48 h,为促进bFGF、VEGF基因和其蛋白表达的最适条件。 OBJECTIVE: To investigate the effects of cobalt chloride (CoCl2) hypobaric preconditioning on gene expression and protein secretion of basic fibroblast growth factor (bFGF) and vascular endothelial growth factor (VEGF) in human umbilical cord mesenchymal stem cells influences. Methods: UC-MSCs were isolated and cultured. The third generation of UC-MSCs were harvested, and the cells were cultured for 72 h with CoCl2 containing 150 μmol / L. Flow cytometry was used to detect the expression of UC-MSCs surface markers (CD34, CD45, CD90, CD105, CD29 and CD49 )Impact. UC-MSCs were divided into 4 groups according to the concentration of CoCl2 (0, 50, 100μmol / L and 150μmol / L), namely control group, low, middle and high CoCl2 groups. h The expression of bFGF and VEGF mRNA was detected by SYBRGREN fluorescence quantitative RT-PCR. The expression of bFGF and VEGF protein was detected by ELISA. Results: Hypoxia in 150μmol / L CoCl2 medium did not change the cell morphology and surface markers. Under the same hypoxia culture time, the expression of bFGF and VEGF increased with the increase of CoCl2 concentration, and reached the peak when CoCl2 was 150 μmol / L (P <0.05). The expression of bFGF and VEGF increased with the increase of hypoxia preconditioning time (P <0.05) when the concentration of CoCl2 was 150 μmol / L for different time (24 h, 48 h and 72 h) <0.05). The expressions of bFGF and VEGF genes were 8.15 ± 0.10 and 16.67 ± 0.17, respectively. The expression of time gene in hypoxic culture was decreased (P <0.05). The changes of protein and gene expression were basically the same. The expression of bFGF and VEGF peaked at 48 h in culture media containing 150 μmol / L CoCl 2 (P <0.05), which were 69.63 ± 7.90 ng / L and 89.55 ± 5.45 ng / L, respectively. Conclusion: The appropriate concentration of CoCl2 (<150μmol / L) has little effect on the biological function of UC-MSCs. CoCl2 can be used to simulate cell hypoxia, hypoxia on UC-MSCs bFGF, VEGF secretion in a time and dose-dependent manner with 150μmol / L CoCl2 culture medium 48h, to promote bFGF, VEGF gene and its protein expression The most suitable conditions.