Yeungnam Univ J Med.  2006 Dec;23(2):182-192. 10.12701/yujm.2006.23.2.182.

Isolation of Endothelial Cells and Smooth Muscle Cells from Rat Aort

Affiliations
  • 1Department of Anatomy, College of Medicine, Yeungnam University, Daegu, Korea. jookim@med.yu.ac.kr

Abstract

BACKGROUND
Atherosclerosis has emerged as the leading cause of death in developed countries. At present, human umbilical vein endothelial cells (HUVEC) are most commonly used for the investigation of Endothelial cells (EC). However, HUVEC are not found in arteries but only in veins. Currently there are many reports on methods used to isolate EC;, most of these methods require special equipment to remove contaminating smooth muscle cells (SMC).
MATERIALS AND METHODS
The method described here may be used to isolate not only ECs but also SMCs;,the approach presented here did not require special equipment. Rat aorta was treated with 2 mg/ml of type II collagenase solution for 45 minutes. The isolated cells from the aorta were incubated in medium G for a week;, only ECs could be separated. After the collagenase treatment, the rest of aorta was cut lengthwise, and left undisturbed to obtain SMCs in the culture dish for 10 days. To verify the purity of the isolated cells, we performed immunofluorescence and evaluated the results with transmission electron microscopy analysis.
RESULTS
The immunofluorescence study demonstrated specific expression of CD31 and alpha-smooth muscle actin in the isolated ECs and SMCs, respectively. Cultured ECs and SMCs showed their own fine structure characteristics.
CONCLUSION
These results suggest that this method for isolating ECs and SMCs may be especially useful for the study of atherosclerosis.

Keyword

Atherosclerosis; Endothelial cell; Smooth muscle cell; CD31; alpha-smooth muscle actin
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