The Cold Preservation of Mice Hepatocytes Induces Apoptosis but Not over Expression of MHC Class I Antigen / 대한이식학회지

ABSTRACT

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.