Chinese Journal of Tissue Engineering Research ›› 2014, Vol. 18 ›› Issue (37): 5961-5966.doi: 10.3969/j.issn.2095-4344.2014.37.010

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Comparison of methods for culturing the mesenchymal stem cells from human umbilical cord blood

Huang Hong-yu1, Liu Guo-ping2, Duan Li2, 3, Chen Yun-fang2, 3, Xiong Jian-yi2, 3, Wang Da-ping2, 3   

  1. 1Shantou University Medical College, Shantou 515000, Guangdong Province, China
    2Second People’s Hospital of Shenzhen, Shenzhen 518035, Guangdong Province, China
    3Shenzhen Key Laboratory of Tissue Engineering, Shenzhen 518035, Guangdong Province, China
  • Revised:2014-07-12 Online:2014-09-03 Published:2014-09-03
  • Contact: Wang Da-ping, M.D., Professor, Chief physician, Doctoral supervisor, Second People’s Hospital of Shenzhen, Shenzhen 518035, Guangdong Province, China; Shenzhen Key Laboratory of Tissue Engineering, Shenzhen 518035, Guangdong Province, China
  • About author:Huang Hong-yu, Studying for master’s degree, Shantou University Medical College, Shantou 515000, Guangdong Province
  • Supported by:

    the Natural Science Foundation of Guangdong Province, No. S2012010008129; the Funds of Science and Technology Commission of Shenzhen Municipality, No. ZDSY20120614154551201, GJHZ20130412153906739, CXZZ20120614160234842, JSGG20140519105550503

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Abstract:

BACKGROUND: The umbilical cord blood is rich of mesenchymal stem cells, which can be used as a new source of seed cells in tissue engineering.
OBJECTIVE: To compare two methods for culturing, expanding and purifying the mesenchymal stem cells in vitro isolated from the human umbilical cord blood.
METHODS: The full-term birth cord blood of 40 cases was collected under sterile conditions with heparin anticoagulation. Ficoll density gradient centrifugation was used to isolate the mononuclear cells from the umbilical cord blood. The cases were randomly divided into two groups according to the different culture media. Twenty cases of umbilical cord blood were cultured in MesenGro human mesenchymal stem cells culture medium (group A), and the remaining 20 cases of umbilical cord blood were cultured in Dulbecco’s modified Eagle’s medium(group B). The occurrence time of fusiform mesenchymal stem cells and cell colony, culture time and the number of primary cells in the two groups were compared. Cells which grew well were selected to detect the surface markers by flow cytometry.
RESULTS AND CONCLUSION: The mean occurrence time of fusiform mesenchymal stem cells and cell colony, culture time and number of primary cells in group A were better than those in group B (P < 0.01). The strong expression of the surface markers of mesenchymal stem cells (CD73 and CD105) was found by flow cytometry, of which the positive rate was 99.1%. No expression of the surface markers of hematopoietic stem cells (CD45 and CD34) was seen, of which the negative rate was 99.3%. The number, morphology, growth rate and culture time of umbilical cord blood mesenchymal stem cells cultured in MesenGro human mesenchymal stem cells culture medium were better than those cultured in Dulbecco’s modified Eagle’s medium. Cells cultured in MesenGro human mesenchymal stem cell culture medium can better express surface markers of 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: fetal blood, mesenchymal stem cells, culture media, cells, cultured

CLC Number: