Chinese Journal of Tissue Engineering Research ›› 2010, Vol. 14 ›› Issue (32): 5952-5956.doi: 10.3969/j.issn.1673-8225.2010.32.014

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Isolation, culture and identification of menstrual blood-derived mesenchymal stem cells

Zhou Yun-fan 1, 2, Yang Bo 2, 3, Hu Xiang 4, 5, Jiao Hong-liang 2, 3, Zhou Chang-hui1, Tian Yi 2, 3, Lei Ning-jing1, Gu Chen-xi6, Xu Yu-ming7, Dai Jian-wu8, Guan Fang-xia1   

  1. 1 Department of Bioengineering, Zhengzhou University, Zhengzhou  450001,  Henan Province, China; 2 Open Laboratory of Clinical Medicine and Key  Subject, H enan Institution of Higher Education, Zhengzhou  450052,  Henan Province, China; 3 Department of Neurosurgery, First Affiliated Hospital, Zhengzhou University, Zhengzhou  450052, Henan Province,  China; 4 Jiangsu Public Technology Service Platform of Stem Cells and Biotherapy, Taizhou  225300, Jiangsu Province, China; 5 Shenzhen Beike Cell Engineering Institute, Shenzhen  518000, Guangdong Province,  China; 6 Department of Clinical Medicine, Zhengzhou University,  Zhengzhou  450001, Henan Province, China; 7 Department of Neurology,  First Affiliated Hospital, Zhengzhou University, Zhengzhou  450052, Henan Province, China; 8Institute of Genetics and Developmental  Biology, Chinese Academy of Sciences, Beijing  100190, China
  • Online:2010-08-06 Published:2010-08-06
  • Contact: Guan Fang-xia, Doctor, Professor, Department of Bioengineering, Zhengzhou University, Zhengzhou 450001, Henan Province, China guanfangxia@ 126.com
  • About author:Zhou Yun-fan★, Studying for master’s degree, Department of Bioengineering, Zhengzhou University, Zhengzhou 450001, Henan Province, China; Open Laboratory of Clinical Medicine and Key Subject, Henan Institution of Higher Education, Zhengzhou 450052, Henan Province, China boat-1986@163.com
  • Supported by:

     the 211 Project-phase Ⅲ of Zhengzhou University- the Basic and Clinical Research of Stem Cells; the Developmental Project of Science and Technology from Department of Science and Technology of Henan Province in 2009, No. 092102310250; the Young Teachers Funded Projects of Henan Province; the Erqi District Society Development Project from Bureau of Science and Technology in Zhengzhou City, No. 20095157; the Public Technology Service Platform of Stem Cells and Biotherapy of Jiangsu Province, No. BM2008146

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Abstract:

BACKGROUND: Generally, the main sources of stem cells are embryonic stem cells and bone marrow stem cells, but there are some limitations of the two kinds of sources, and researchers strive to find a new type of stem cells in order to overcome the potential for teratoma, the lack of sample sources, ethics controversies and so on.
OBJECTIVE: To isolate, culture and identify menstrual blood-derived mesenchymal stem cells (MMCs).
METHODS: MMCs were separated by Ficoll density gradient centrifugation. Cell morphology and growth characteristics were observed. Flow cytometry analysis was used to identify expression of CD29, CD44, CD34, CD45, HLA-ABC and HLA-DR surface markers in MMCs. Immunocytochemical method was utilized to detect nestin-positive cells in MMCs.
RESULTS AND CONCLUSION: MMCs were successfully separated from menstrual blood by Ficoll density gradient centrifugation. About 2 weeks later, primary cells reached 80% to 90% confluence; cells were spiral-shaped, mesh and radial. After passage to a stable growth, cells could be seen as the leading fiber-like cell morphology. Results from flow cytometry showed that MMCs were highly positive for CD29, positive for CD44, low positive for HLA-ABC, but negative for CD34, CD45 and HLA-DR. Staining showed nestin expression of antigen in MMCs was about (10.35±0.51)%. Results have suggested that MMCs express MSCs markers, and only have low immunogenicity. MMCs express nestin antigen and provide some theoretical basis for MMCs in the nervous system application.

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