中国组织工程研究Issue(45)：7268-7273,6.DOI:10.3969/j.issn.2095-4344.2015.45.010
氯化钴模拟低氧环境下人脐带间充质干细胞增殖及基因蛋白的表达
Effect of cobalt chloride-induced hypoxia on proliferation of human umbilical cord-derived mesenchymal stem cells and related gene and protein expressions
摘要
Abstract
BACKGROUND:Cobalt chloride (CoCl2) may promote the proliferation of human umbilical cord-derived mesenchymal stem cels (hUC-MSCs) in a time- and concentration-dependent manner, and meanwhile, CoCl2 can regulate the expression of genes and proteins in hUC-MSCs. OBJECTIVE:To explore the effects of CoCl2 induced-hypoxia on the proliferation of hUC-MSCs and gene and protein expressions in hUC-MSCs, thereby establishing an effective method for MSCs culture and amplificationin vitro. METHODS: hUC-MSCs were extracted using tissue explant method. Under hypoxia conditions induced by CoCl2 (0, 100, 150, 200, 250 μmol/L) for different periods (0, 1, 2, 3, 4 days), flow cytometry was used to identify cel surface-associated antigens; cel counting kit-8 was used to detect cel proliferation; RT-PCR was used to determine levels of hypoxia inducible factor-1α, inducible nitric oxide synthase, stromal cel-derived factor-1, interleukin-6, transforming growth factor-β mRNA; western blot assay was used to detect protein expression of hypoxia inducible factor-1α. RESULTS AND CONCLUSION:The cels were positive for CD29, CD73, CD90, CD105, while negative for CD31, CD14, CD34, CD45, CD11b, HLA-DR. Moreover, the antigen expression was not affected by CoCl2 induced-hypoxia. CoCl2 induced-chemical hypoxia could promote the proliferation of hUC-MSCs in a time- and concentration-dependent manner. RT-PCR results showed thatunder hypoxia, hypoxia inducible factor 1α, inducible nitric oxide synthase and stromal cel-derived factor-1 mRNA expressions were significantly up-regulated, but interleukin-6 and transforming growth factor-β mRNA expressions were down-regulated significantly (P < 0.05). Additionaly, the protein expression of hypoxia inducible factor 1α was increased under hypoxia conditions. These findings indicate that CoCl2 induced-hypoxia environment may promote the proliferation of hUC-MSCs and the optimal concentration of CoCl2 is 200μmol/L. However, a higher concentration of CoCl2 (≥ 250μmol/L) inhibits the proliferation of hUC-MSCs, and the mechanism may be related to the increase of hypoxia inducible factor-1α at protein and mRNA levels.关键词
干细胞/脐带脐血干细胞/人脐带间充质干细胞/低氧/氯化钴/增殖/基因表达/缺氧诱导因子1αKey words
Umbilical Cord/Mesenchymal Stem Cels/Cel Hypoxia/Cel Proliferation/Hypoxia-Inducible Factor 1/Tissue Engineering分类
医药卫生引用本文复制引用
韩潇,白海,尹娇娇,杨柯,韩燕霞,欧剑锋,王存邦..氯化钴模拟低氧环境下人脐带间充质干细胞增殖及基因蛋白的表达[J].中国组织工程研究,2015,(45):7268-7273,6.基金项目
甘肃省科技重大专项(1102FKDA005)Funding:the Major Scientific Project of Gansu Province, No.1102FKDA005 （1102FKDA005）
