- Isolation of Endothelial Cells and Smooth Muscle Cells from Rat Aort.
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Young Eun Yun, In Hwan Song, Eon Ki Sung, Joo Young Kim
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Yeungnam Univ J Med. 2006;23(2):182-192. Published online December 31, 2006
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DOI: https://doi.org/10.12701/yujm.2006.23.2.182
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 Abstract
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- 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.
- Transition of Marker Enzymes of Rat Hepatocyte Organelles in Culture.
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In Hwan Song, Joo Yung Kim, Eon Ki Sung, Yung Chang Lee
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Yeungnam Univ J Med. 1989;6(2):133-140. Published online December 31, 1989
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DOI: https://doi.org/10.12701/yujm.1989.6.2.133
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Abstract
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- To investigate recovery, growth, and activity of hepatocyte in primary culture after cell separation, the authors followed up the marker enzyme activities of golgi complex, mitochondria and biologic membrane. Thiamine pyrophosphatase, the marker enzyme of golgi complex, activity approached the level of long term culture at 4th day. Succinate dehydrogenase, the marker enzyme of mitochondria, activity decreased with time, then it maintained constant level after 4th day. Alkaline phosphatase, the marker enzyme of biological membrane, activity increased from 3rd day, and after 5th day it showed strong reaction. These data suggested that hepatocytes were stabilized and recovered normal activity 4 day after cell separation. But the main secretory function was speculated to be reduced in culture.
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Yeungnam University College of Medicine
