[Effect of the composition of liver perfusion media on functional properties of isolated hepatocytes and effectiveness of the isolation method].
Yield, viability and respiratory activity of hepatocytes isolated from rat after liver perfusion by media with various composition have been studied. Collagenase, EDTA or citrate were used as disaggregative agents, as well as sucrose or NaCl as a main osmotic component. The yield of intact cells after perfusion by different media decreased in the following order, collagenase -NaCl-->EDTA-sucrose-->citrate-sucrose-->EDTA-NaCl. The rate of endogenous respiration in hepatocytes isolated by non-enzymatic methods with EDTA did not depend on the nature of main osmotic component and was by 40% lower than in the cells isolated by the enzymatic method, however uncoupler-stimulated respiration changed significantly less. Replacement of EDTA by citrate increased the rate of endogenous respiration and particularly the uncoupler-stimulated respiration. The results show that the nature of disaggregative agent and main osmotic component has the influence on efficiency of cell isolation method as well as metabolic state of isolated hepatocytes. Sucrose leads to the cell shrinkage and have additional to Ca(2+)-chelator desaggregative effect, citrate plays two functions both as Ca(2+)-chelator and as mitochondrial substrate.
Petrenko
A Iu
AIu
Govorukha
T P
TP
D000818:Animals; D002469:Cell Separation / Q000592:standards*; D003470:Culture Media; D008099:Liver / Q000166:cytology / Q000378:metabolism / Q000502:physiology*; D009995:Osmosis; D010100:Oxygen / Q000378:metabolism; D010477:Perfusion; D051381:Rats; D017208:Rats, Wistar
D003470:Culture Media; D010100:Oxygen