Chinese Journal of Tissue Engineering Research ›› 2010, Vol. 14 ›› Issue (50): 9366-.doi: 10.3969/j.issn.1673-8225.2010.50.014

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Magnetic beads isolation and primary culture of mouse pulmonary microvascular endothelial cells in vitro

Sun Zhen-zhen1, Cai Zai-long2, Zhu Ke-ming1, Deng Xiao-ming1   

  1. 1 Department of Anesthesiology, 2 Clinical Experimental Center, Changhai Hospital, Second Military Medical University of  Chinese PLA, Shanghai  200433,  China
  • Online:2010-12-10 Published:2010-12-10
  • Contact: Zhu Ke-ming, Professor, Chief physician, Department of Anesthesiology, Changhai Hospital, Second Military Medical University of Chinese PLA, Shanghai 200433, China kmzhu_md@yahoo.com.cn Deng Xiao-ming, Professor, Chief physician, Department of Anesthesiology, Changhai Hospital, Second Military Medical University of Chinese PLA, Shanghai 200433, China xmdeng@anesthesia.org.cn
  • About author:Sun Zhen-zhen☆, Studying for doctorate, Attending physician, Department of Anesthesiology, Changhai Hospital, Second Military Medical University of Chinese PLA, Shanghai 200433, China zhenzhensun2008@hotmail.com
  • Supported by:

    the National Natural Science Foundation of China, No. 30872459*

     

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Abstract:

BACKGROUND: Lung function depends on the activity of pulmonary microvascular endothelial cells (PMVECs). It is a very important cell model in related researches, while the cells which are isolated by the tissue block culture adhesion method that are currently widely used in China have a major disadvantage of being usually mixed with other cells.
OBJECTIVE: To establish an accurate and effective technique and methods of isolation and culture mouse PMVECs in vitro.
METHODS: PMVECs were isolated and purified by collagenase I digesting and magnetic beads separating,and adherent cultured in vitro. The growth curve of cultured PMVECs was measured by CCK-8, the cell morphology and ultrastructure was observed by microscopes and its phenotype was identified by flow cytometry.
RESULTS AND CONCLUSION: The cells grew as a monolayer in culture and exhibited the well-described cobblestone morphology, contained a lot of endothelial cell specific rhabditiform organelles Weibel-Palade bodies, expressed endothelial cell specific surface marker CD105 relatively stably, and rarely expressed lymphatic endothelial cell specific surface marker VEGFR-3. The PMVECs isolated successfully by immunoglobulin magnetic beads are of high purity and rapid multiplication and retain certain of their functional differences, including constitution and antigen molecule expression.

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