Chinese Journal of Tissue Engineering Research ›› 2012, Vol. 16 ›› Issue (1): 27-30.doi: 10.3969/j.issn.1673-8225.2012.01.005

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Isolation and charcaterization of human bone marrow mesenchymal stem cells using a whole bone marrow adherent culture technique

Zhang Hao1, 2, Zhang Bin2, Cheng Mei1, Tao Yan-ling1, Hu Jiang-wei2, Xu Man2, Chen Hu2   

  1. 1Harbin Institute of Hematology Oncology, the First Hospital of Harbin, Harbin  150010, Heilongjiang Province, China; 2Hematopoietic Stem Cell Transplantation Center, Affiliated Hospital of Academy of Military Medical Science of Chinese PLA, Beijing  100071, China The First Hospital of Harbin and the Affiliated Hospital of Academy of Military Medical Science of Chinese PLA were co-considered as the first units.
  • Received:2011-10-25 Revised:2011-11-24 Online:2012-01-01 Published:2012-01-01
  • Contact: Correspondence to: Chen Hu, Doctor, Chief physician, Hematopoietic Stem Cell Transplantation Center, Affiliated Hospital of Academy of Military Medical Science of Chinese PLA, Beijing 100071, Chinachenhu217@yahoo.com.cn
  • About author:Zhang Hao☆, Doctor, Associate chief physician, Harbin Institute of Hematology Oncology, the First Hospital of Harbin, Harbin 150010, Heilongjiang Province, China; Hematopoietic Stem Cell Transplantation Center, Affiliated Hospital of Academy of Military Medical Science of Chinese PLA, Beijing 100071, Chinagx-zhanghao@126.com Zhang Bin☆, Doctor, Associate chief physician, Hematopoietic Stem Cell Transplantation Center, Affiliated Hospital of Academy of Military Medical Science of Chinese PLA, Beijing 100071, Chinazb307ctc@163.com

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Abstract:

Abstract
BACKGROUND: Bone marrow mesenchymal stem cells (BMSCs) are widely used in cell therapy and tissue regeneration, so their isolation and culture are a key technique for research and clinical application.
OBJECTIVE: To isolate the BMSCs by whole bone marrow adherent culture technique and to establish an easy and effective method to isolate and culture BMSCs.
METHODS: The BMSCs were isolated by whole bone marrow adherent culture technique, and purified and amplified by passage. BMSCs were induced to differentiate into adipocytes, osteocytes and chondrocytes in special differentiation condition.   
RESULTS AND CONCLUTION: The purity of the BMSCs isolated by whole bone marrow adherent culture technique was more than 90%. The adherent cells displayed an abundant presence of CD73, CD90, CD105 and absence of Hematopoietic cell phenotype CD34, CD45 and HLA-DR which were detected by flow cytometry. The cell doubling time was (24.04±0.49) hours; Cell cycle showed that there was percentage of stem cells as G0 /G1 71.63% and S+G2+M 28.37% respectively. The induced and differentiated results showed than BMSCs could be differentiated into adipocytes, osteocytes and chondrocytes. Based on these findings, the whole bone marrow adherent culture technique is a better method to obtain MSCs from human bone marrow.

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