Chinese Journal of Tissue Engineering Research ›› 2011, Vol. 15 ›› Issue (1): 12-16.

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Isolation and identification of rabbit bone marrow mesenchymal stem cells by low permeability combined with natural sedimentation

Lin Chun-bo1, Yang Yuan1, Chen Wei-ping2   

  1. 1Guangxi Bone Trauma Hospital, Nanning  530012, Guangxi Zhuang Autonomous Region, China
    2Guangxi Medical University, Nanning  530021, Guangxi Zhuang Autonomous Region, China
  • Received:2010-07-25 Revised:2010-10-14 Online:2011-01-01 Published:2011-01-01
  • Contact: Yang Yuan, Professor, Guangxi Bone Trauma Hospital, Nanning 530012, Guangxi Zhuang Autonomous Region, China; Chen Wei-ping, Professor, Guangxi Medical University, Nanning 530021, Guangxi Zhuang Autonomous Region, China
  • About author:Lin Chun-bo★, Master, Physician, Guangxi Bone Trauma Hospital, Nanning 530012, Guangxi Zhuang Autonomous Region, China linchunbo1983@163.com
  • Supported by:

    Grants of Department of Science and Technology of Guangxi Zhuang Autonomous Region of China, No. Guikegong 0816004-15; Guikezi 0728077; Guikezi 0832157; Guikegong 0592007-1G

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Abstract:

BACKGROUND: The number of bone marrow mesenchymal stem cells (BMSCs) was few. 100 000 bone marrow karyocytes contain 1 BMSC. How to successfully isolate and identify BMSCs is a key in the study.
OBJECTIVE: To establish a perfect culture system in vitro of BMSCs; BMSCs were identified from cell morphology, phenotype and functional aspects.
METHODS: Low permeability principle was used to remove mixed cell, and combined with the natural sedimentation to isolate BMSCs. Their morphological characteristics were observed by light microscopy. The cell phenotype of BMSCs was determined by flow cytometry. BMSCs were induced into osteoblasts and adipocytes. Cell morphology was observed and alkaline phosphatase (AKP), Von Kossa, collagen typeⅠ, oil red O detection were performed.
RESULTS AND CONCLUSION: BMSCs were identified as phenotype: CD29(+), CD45(-). Following being induced into osteoblasts, AKP, mineralized nodules and collagen type Ⅰ staining were detected positively; to adipocytes, the oil red O staining was detected positively. Results suggest that isolated BMSCs are non-hematopoietic stem cells with fibroblast-like cell morphology. These cells had the functions of differentiating into osteoblasts and adipocytes.

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