Chinese Journal of Tissue Engineering Research ›› 2014, Vol. 18 ›› Issue (19): 3042-3047.doi: 10.3969/j.issn.2095-4344.2014.19.015

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Culture methods for primary human umbilical cord mesenchymal stem cells

Wang Fei1, Zhou Hong1, Guo Yu-cheng1, Su Xiao-xia1, Li Ang1, Zhao Xiao-peng2   

  1. 1Department of Orthodontics, Hospital of Dentistry, Xi’an Jiaotong University, Xi’an 710004, Shaanxi Province, China
    2The Hospital of No. 205 Research Institute of Weapons Industry in China, Xi’an 710065, Shaanxi Province, China
  • Revised:2014-03-08 Online:2014-05-07 Published:2014-05-07
  • Contact: Zhou Hong, Professor, Department of Orthodontics, Hospital of Dentistry, Xi’an Jiaotong University, Xi’an 710004, Shaanxi Province, China
  • About author:Wang Fei, M.D., Attending physician, Department of Orthodontics, Hospital of Dentistry, Xi’an Jiaotong University, Xi’an 710004, Shaanxi Province, China
  • Supported by:

    the Natural Science Foundation of Shaanxi Province, No. 2014JM4146; the “13115” Scientific Innovation Engineering Plan of Shaanxi Province, No. 2008ZDKG-65

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Abstract:

BACKGROUND: How to get a lot of stable and dynamic human umbilical cord mesenchymal stem cells is still a difficulty.
OBJECTIVE: To investigate the best culture way of human umbilical cord mesenchymal stem cells by comparing tissue explant, enzyme digestion, enzymolysis methods.
METHODS: Ten fresh umbilical cords were isolated to human umbilical cord mesenchymal stem cells by using tissue explant, enzyme digestion, and enzymolysis methods, respectively. Hereafter, we compared cells harvested using three culture methods in terms of the time for the primary cells to creep, successful rates of cell-culturing, and curves of cell growth. Surface markers of cells were detected by flow cytometry and multiple differentiations of cells were detected.
RESULTS AND CONCLUSION: The time for the primary cells to creep out in the enzymolysis and enzyme digestion groups had no significant difference, but both of them were significantly shorter than that in the tissue explant group (P < 0.01). The successful rate of primary cell culture in the enzymolysis group was significantly higher than that in the other two groups. The cell proliferations of three groups had no significant difference. The cell surface markers and differentiation ability of the third generation of human umbilical cord mesenchymal stem cells cultured by enzymolysis method met the characteristics of mesenchymal stem cells. In conclusion, the enzymolysis method can shorten the time for primary human umbilical cord mesenchymal stem cells to creep out and increase the successful rates of primary human umbilical cord mesenchymal stem cells.


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


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Key words: stem cells, mesenchymal stem cells, umbilical cord, cells, cultured

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