Chinese Journal of Tissue Engineering Research ›› 2010, Vol. 14 ›› Issue (10): 1739-1742.doi: 10.3969/j.issn.1673-8225.2010.10.006

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Isolation, cultivation and reproductive activity of human umbilical cord blood mesenchymal stem cells

Yan Man1, Yang Yi-yong2, Qin Shu-jian2, Zheng De-yu2   

  1. 1Department of Hand Surgery, First Affiliated Hospital, Liaoning Medical University, Jinzhou   121001, Liaoning Province, China;
    2Department of Anatomy, Liaoning Medical University, Jinzhou   121001, Liaoning Province, China
  • Online:2010-03-05 Published:2010-03-05
  • Contact: Zheng De-yu, Doctor, Associate professor, Department of Anatomy, Liaoning Medical University, Jinzhou 121001, Liaoning Province, China zheng_deyu2000@yahoo.com.cn
  • About author:Yan Man, Nurse-in-charge, Department of Hand Surgery, First Affiliated Hospital, Liaoning Medical University, Jinzhou 121001, Liaoning Province, China
  • Supported by:

    the Youth Foundation of Department of Education of Liaoning Province, No. 05L140*

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Abstract:

BACKGROUND: With increased age, bone marrow mesenchymal stem cells (BMSCs) are influenced with regard to quantity and quality, which will induce great damages to the donors. Many studies have focused on seeking substitute MSC source. In contrast, it remains controversial whether umbilical cord blood contains MSCs.

OBJECTIVE: To isolate MSCs from human umbilical cord blood, and to detect their biological properties.

METHODS: Umbilical cord blood samples were sterilely isolated using Percoll density gradient centrifugation to harvest intermediate layer cells. DMEM medium containing fetal bovine serum, penicillin, streptomycin and L-glutamine was added. Following several adherences and purification, the floating cells were discarded. Thus, many adherent cells with a confluence were collected. When cells were 60%-80% confluent, cells were digested by trypsin for subculture. Cells at passages, 1, 5 and 9 were obtained and their morphological changes were observed. Cell surface antigens were measured using flow cytometry. Growth curves were drawn, and cell viability was determined utilizing MTT.

RESULTS AND CONCLUSION: Isolated umbilical cord blood MSCs presented an even size, showing spindle or star-shape fibroblasts-like cells. Umbilical cord blood MSCs at 1, 5, 9 passages were greatly positive for CD29, CD105 and CD166, but weakly positive for CD34 and CD45. Following 5 days of incubation, cells entered logarithmic growth phase. The number was decreased at day 9. Population doubling time was (53.5±8.32) hours. Cells grew well. Cells at 1-7 passages showed similar viability (P > 0.05). Till passage 9, cell proliferation viability was decreased, but no significant difference was determined (P > 0.05). Results verified that MSCs can be successfully isolated from umbilical cord blood in vitro. Cells at passages 1-9 presented a good reproductive activity.

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