This study deals with isolation of @entity35 hepatocytes by a non-enzymatic method and the separation of intact and damaged cells in @entity969 medium. Low speed centrifugation in isotonic @entity969 medium of a hepatocyte suspension obtained by mechanical desaggregation of liver pre-perfused with @entity1098 solution results in the formation of a cell pellet which contains two different layers. A darker layer contains hepatocytes with intact plasma membranes. Their respiratory activity and xenobiotic metabolism are close to those of the cells isolated by collagenase perfusion. The study of distribution of lipophilic cation @entity36958 ( @entity8331 ) indicates a predominantly mitochondrial localization of @entity8331 in the intact cells following non-enzymatic and collagenase isolation. Hepatocytes in the upper layer have damaged plasma membranes. As a result they lose the potential to accumulate @entity8331 , and have low rates of endogenous respiration and biotransformation activity. Addition of exogenous @entity1110 restores the capability to metabolize xenobiotics. Washing and incubation of these hepaticytes in an intracellular type medium results in restoration of uncoupler-stimulated @entity26 consumption and generation of membrane potential in the presence of a @entity4588 substrate. These properties are close to those of hepatocytes permeabilized by @entity11040 treatment. Thus, the procedure allows the simultaneous isolation of both intact and permeabilized hepatocytes with functionally active intracellular structures without the use of relatively expensive chemicals such as collagenase and Percoll.

Separation of intact and damaged hepatocytes in XXXX following non-enzymatic liver perfusion.

@entity8331 :: ('CHEBI:52279', 'Chemical') :: ['TPP(+)'] @entity11040 :: ('MESH:D004072', 'Chemical') :: ['digitonin'] @entity4588 :: ('MESH:D019802', 'Chemical') :: ['succinate'] @entity26 :: ('MESH:D010100', 'Chemical') :: ['oxygen'] @entity1110 :: ('MESH:D009249', 'Chemical') :: ['NADPH'] @entity969 :: ('MESH:D013395', 'Chemical') :: ['sucrose'] @entity36958 :: ('MESH:C013289', 'Chemical') :: ['tetraphenylphosphonium'] @entity35 :: ('10116', 'Species') :: ['rat'] @entity1098 :: ('MESH:D004492', 'Chemical') :: ['EDTA']
@entity969 :: (MESH:D013395,Chemical) :: ['sucrose']
