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KMID : 0361120010150010058
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Korean Journal of Transplantation
2001 Volume.15 No. 1 p.58 ~ p.66
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The Cold Preservation of Mice Hepatocytes Induces Apoptosis but Not over Expression of MHC Class I Antigen
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이은아/Lee EA
박선주/최대화/고병균/남창우/나양원/김규열/박건춘/이영주/박광민/한덕종/이승규/최석영/김병삼/조홍래/Park SJ/Choi DH/Ko BK/Nam CW/Nah YW/Kim GY/Park KC/Lee YJ/Park KM/Han DJ/Lee SG/Choe SY/Kim BS/Cho HR
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Abstract
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Hepatocyte transplantation is a potential treatment modality for liver diseased patients. Purified hepatocytes stimulates allospecific cytotoxicity by expressing the MHC class I antigen. Also, during cold preservation, hepatocytes are damaged by lipid peroxidation with oxygen free radicals, which may induce apoptosis on cold preserved hepatocyte. For measuring the degree of antigenicity on cold- preserved mice hepatocytes with UW solution, we studied the expression of MHC class I antigen in various time period by FACS and RT-PCR. For analysis of apoptotic hepatocyte death, we studied morphological changes and DNA fragmentation. We used flow cytometry techniques with rhodamine 123,3,3’-dihexiloxadicarbocyanine (DiOC6 (3)) and propidium iodide (PI). DiOC6 (3) is mitochondrial probe to measure the mitochondrial transmembrane potential that drops early in apoptosis. The percentage of cells undergoing chromatinolysis (subdiploid cells) was determined by ethanol fixation followed by RNA digestion and PI staining. The cold preserved hepatocytes expressed MHC class I constitutively, but revealed no significant differences among various preservation period. However, apoptosis of hepatocytes occured progressively during cold preservation. These results provides that the cold preservation of mice hepatocyte induces apoptosis with involvement of an oxidative process, but does not stimulate over expression of MHC class I antigen.
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