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KMID : 0363720090420020093
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Korean Journal of Anatomy
2009 Volume.42 No. 2 p.93 ~ p.104
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An Effective Isolation of the Vascular Endothelial and Smooth Muscle Cells from the Mouse Aorta
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Kim Kyung-Gon
Sung Eon-Gi
Kim Joo-Young
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Abstract
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The mechanism of the disease such as artherosclerosis is easily elucidated by the comparison among cells isolated from each aorta of knockout mouse and wild type mouse, respectively. This study was aimed at effectively harvesting the endothelial and smooth muscle cells from 4~6 weeks old wild type C57BL/6J mouse aorta. The tunica adventitia was completely removed to get the aortic tissues only consisting of the tunica intima and the tunica media under the stereoscope. These aortic tissues were treated with type I collagenase or type II collagenase solution, respectively, and then the endothelial or smooth muscle cell was isolated. CD31 marker of the endothelial cell and α-smooth muscle actin marker of the smooth muscle cell were identified with confocal microscope. The percentages of the labelled cells by each marker represented the extent of purification of endothelial or smooth muscle cells, respectively, for harvested cells according to the collagenase solutions. 70~80% of culture vessel was covered with the endothelial cells 10 days after the treatment of the type I collagenase solution, while 40~50% of culture vessel covering with the cells after the treatment of the type II collagenase solution. 70~80% of culture vessel was covered with the smooth muscle cell regardless of the type of the collagenase solution on the 13th day. Percentages of the CD31 positive cells after the treatment with the type I or the type II collagenase solution was 91.1±2.865%** and 86.4±3.641%, respectively (**p ?0.05, n=5). Percentages of the α-smooth muscle actin labelled cells after the treatment with the type I or the type II collagenase solution were 87.9±5.713% and 86.6±4.778%, respectively, and these values were not significantly different. Taken together, the aortic tissues using the type I collagenase solution comparing with using the type II collagenase solution were much more effective in the isolation of the endothelial cells.
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KEYWORD
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Endothelial cells, Smooth muscle cells, Mouse aorta, Type I collagenase solution, Type II collagenase solution
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