Chinese Journal of Tissue Engineering Research ›› 2012, Vol. 16 ›› Issue (36): 6762-6766.doi: 10.3969/j.issn.2095-4344.2012.36.020

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Whole bone marrow cells culture for expanding mouse endothelial progenitor cells

Liu Jun-feng1, Du Zhong-dong1, Chen Zhi1, Guan Yun-qian2, Li Shen-tao3   

  1. 1Department of Cardiology, Beijing Children’s Hospital of Capital Medical University, Beijing 100045, China;
    2Cell Biology Laboratory Affiliated Xuanwu Hospital of Capital Medical University, Beijing 100053, China;
    3School of Basic Medical, Capital Medical University, Beijing 100069, China
  • Received:2011-11-28 Revised:2011-12-15 Online:2012-09-02 Published:2012-09-02
  • Contact: Du Zhong-dong, Professor, Chief physician, Doctoral supervisor, Department of Cardiology, Beijing Children’s Hospital of Capital Medical University, Beijing 100045, China duzhongdong@vip.Sohu.com
  • About author:Liu Jun-feng☆, Studying for doctorate, Attending physician, Department of Cardiology, Beijing Children’s Hospital of Capital Medical University, Beijing 100045, China liuboxuan@126.com

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Abstract:

BACKGROUND: The protocols of culturing and expanding endothelial progenitor cells (EPCs) through isolating bone marrow mononuclear cells are complex in small laboratory animals.
OBJECTIVE: To explore the feasibility of whole bone marrow cells culture for expanding EPCs.
METHODS: EPCs were separated from C57BL/6 mice by whole bone marrow cells culture, and the EPCs were detected with DiL-acLDL and FITC-UEA-1 double fluorescent staining after cultured for 7 days, and the expression of CD34 and FLK-1 was detected by flow cytometry. Furthermore, the functions of EPCs in vitro, including visualization, proliferation, adhesion and migration were assessed. The bone marrow mononuclear cells culture was used as control.
RESULTS AND CONCLUSION: The colony of early EPCs could be seen after cultured for 2 days with whole bone marrow cells culture method in vitro. On day 7, a large number of short spindle-like cells appeared, and exhibited the ability of binding with FITC-UEA-1 and phagocytosis of Dil-acLDL. The EPCs could form vessel like structure when cultured on matrigel. After cultured for about 2 weeks, the late EPCs colonies appeared and proliferated rapidly, in typical cobblestone morphology and could be passaged and cultured in vitro. There was no significant difference between two groups in the number of cells, the expression of CD34 and FLK-1 on the cell surface, in vitro adhesion, proliferation, migration abilities and the time for the colony appearance (P > 0.05). The method of whole bone marrow cells culture in vitro could select and expand EPCs effectively. The procedure is more simple, and especially suitable for studying EPCs functions in the small laboratory animals.

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