Chinese Journal of Tissue Engineering Research ›› 2015, Vol. 19 ›› Issue (45): 7268-7273.doi: 10.3969/j.issn.2095-4344.2015.45.010

Previous Articles     Next Articles

Effect of cobalt chloride-induced hypoxia on proliferation of human umbilical cord-derived mesenchymal stem cells and related gene and protein expressions

Han Xiao1, 2, Bai Hai1, Yin Jiao-jiao1, Yang Ke1, Han Yan-xia1, Ou Jian-feng1, Wang Cun-bang1   

  1. 1Department of Hematology, Center for Hematologic Diseases of Chinese PLA/Lanzhou Military Area General Hospital, Lanzhou 730050, Gansu Province, China; 2Department of Hematology, Second Hospital of Lanzhou University, Lanzhou 730030, Gansu Province, China
  • Received:2015-09-29 Online:2015-11-05 Published:2015-11-05
  • Contact: Bai Hai, M.D., Chief physician, Master’s supervisor, Department of Hematology, Center for Hematologic Diseases of Chinese PLA/Lanzhou Military Area General Hospital, Lanzhou 730050, Gansu Province, China
  • About author:Han Xiao, Studying for master’s degree, Department of Hematology, Center for Hematologic Diseases of Chinese PLA/Lanzhou Military Area General Hospital, Lanzhou 730050, Gansu Province, China; Department of Hematology, Second Hospital of Lanzhou University, Lanzhou 730030, Gansu Province, China
  • Supported by:

    the Major Scientific Project of Gansu Province, No. 1102FKDA005

Abstract:

BACKGROUND: Cobalt chloride (CoCl2) may promote the proliferation of human umbilical cord-derived mesenchymal stem cells (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 amplification in 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 cell surface-associated antigens; cell counting kit-8 was used to detect cell proliferation; RT-PCR was used to 
determine levels of hypoxia inducible factor-1α, inducible nitric oxide synthase, stromal cell-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 cells 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 that under hypoxia, hypoxia inducible factor 1α, inducible nitric oxide synthase and stromal cell-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). Additionally, 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 200 μmol/L. However, a higher concentration of CoCl2 (≥ 250 μmol/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.
中国组织工程研究杂志出版内容重点:干细胞;骨髓干细胞;造血干细胞;脂肪干细胞;肿瘤干细胞;胚胎干细胞;脐带脐血干细胞;干细胞诱导;干细胞分化;组织工程

Key words: Umbilical Cord, Mesenchymal Stem Cells, Cell Hypoxia, Cell Proliferation, Hypoxia-Inducible Factor 1, Tissue Engineering