Chinese Journal of Tissue Engineering Research ›› 2015, Vol. 19 ›› Issue (10): 1477-1481.doi: 10.3969/j.issn.2095-4344.2015.10.001

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Improved primary culture system increases the yield of human umbilical cord mesenchymal stem cells

Gao Yan-lin1, 2, Zhang Ning-kun2, Chen Hou-liang2, Gao Lian-ru2, Zhu Zhi-ming2   

  1. 1Clinical College of Navy Medicine, Anhui Medical University, Beijing 100048, China; 2Navy General Hospital, Beijing 100048, China
  • Online:2015-03-05 Published:2015-03-05
  • Contact: Zhu Zhi-ming, Master, Chief physician, Navy General Hospital, Beijing 100048, China
  • About author:Gao Yan-lin, Studying for master’s degree, Physician, Clinical College of Navy Medicine, Anhui Medical University, Beijing 100048, China; Navy General Hospital, Beijing 100048, China
  • Supported by:

    the National Natural Science Foundation of China, No. 81170094

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Abstract:

BACKGROUND: At present, a lot of research about culture methods for umbilical cord mesenchymal stem cells, but not for the waste of primary system.

OBJECTIVE: To explore the best culture method of human umbilical cord mesenchymal stem cells in vitro.
METHODS: Human umbilical cord mesenchymal stem cells were prepared by tissue explants method, recorded as initial culture group. The centrifugal fluid and tissue of the primary culture flask were centrifuged and divided into three groups for secondary culture: tissue group, mixed group and pure liquid group. Cell morphology, time for cell acquisition, and yield of primary cells in the four groups were observed; the cell growth curve was analyzed by MTT assay; and cell cycle and phenotype were detected by flow cytometry.
RESULTS AND CONCLUSION: The average time for cell acquisition in the initial culture group, tissue group, mixed group and pure liquid group were (15.00±0.45), (7.0±0.3), (8.00±0.25) and (8.00±0.25) days, respectively. The number of cells at first generation was (4.0±0.5)×105, (9.0±0.55)×105, (15.0±0.2)×105 and (7.0±0.33)×105 per T75 culture bottle, respectively. Under the inverted microscope, cells in the four groups were fusiform-like adherent cells, which were in parallel or circinate arrangement. Growth curve, proliferative activity, surface markers of the four groups had no significant differences. The human umbilical cord mesenchymal stem cells can be obtained rapidly and largely through the secondary culture to the primary culture system.
1库：

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: Umbilical Cord, Mesenchymal Stem Cells, Cells, Cultured

CLC Number: