Chinese Journal of Tissue Engineering Research ›› 2012, Vol. 16 ›› Issue (45): 8435-8439.doi: 10.3969/j.issn.2095-4344.2012.45.014

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In vitro isolation, cultivation and identification of ecto-mesenchymal stem cells

Li Bing, Fei Zhou, Hu Shi-jie, Lin Wei, Li Xia, Hu Xue-an, Wang Bing   

  1. Department of Neurosurgery, Xijing Hospital, Fourth Military Medical University, Xi’an 710032, Shaanxi Province, China
  • Received:2012-01-19 Revised:2012-04-03 Online:2012-11-04 Published:2012-11-04
  • Contact: Fei Zhou, Doctor, Professor, Chief physician, Doctoral supervisor, Department of Neurosurgery, Xijing Hospital, Fourth Military Medical University, Xi’an 710032, Shaanxi Province, China feizhou@fmmu.edu.cn
  • About author:Li Bing☆, Doctor, Associate professor, Associate chief physician, Department of Neurosurgery, Xijing Hospital, Fourth Military Medical University, Xi’an 710032, Shaanxi Province, China libingtg1968@163.com

Abstract:

BACKGROUND: Ecto-mesenchymal stem cells are not only a new kind of seed cells in tissue engineering research, but also an innovative therapeutic strategy for tumors in clinical nervous system.
OBJECTIVE: To observe morphology and ultrastructure, identify nerve stem cell marker and test the ability of growth and the proliferation of in vitro isolated ecto-mesenchymal stem cells.
METHODS: Ecto-mesenchymal stem cells were enzymatically isolated from the first branchial arch of rat embryo and maintained in an undifferentiated state with beads coated low affinity nerve growth factor receptor antibody. Then they were purified by magnetic activated cell sorting.
RESULTS AND CONCLUSION: Ecto-mesenchymal stem cells after purification by magnetic activated cell sorting grew like swirls and were fibroblast-like and long spindle-shaped. The ultrastructure of ecto-mesenchymal stem cells was characterized by mesenchymal-like cells with irregular shape, undifferentiation and high activity of proliferation. Immunohistochemical staining manifested that ecto-mesenchymal stem cells expressed human natural killer cells-1, Vimentin and S-100 but not cytokeratin. Ecto-mesenchymal stem cells grew into the exponential phase after cultured for 3 days, and reached to plateau phase after cultured for 6 days. The growth curve was “S” like and the doubling time was 40.28 hours. The evidence suggested that the cells in vitro cultured and purified by magnetic activated cell sorting were undifferentiated ecto-mesenchymal stem cells with stem cell characteristics.

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