AJP - Cell Physiology, Vol 241, Issue 1 3-C8, Copyright © 1981 by American Physiological Society
Deleterious effects of calcium deprivation on freshly isolated hepatocytes
J. W. Edmondson and N. U. Bang
Biochemical and morphological properties of rat hepatic parenchymal cells
isolated without calcium were compared to cells isolated by adding calcium
to the isolation medium at the time of addition of collagenase. Calcium
contents of the two cell preparations were 4.5 +/- 0.3 and 10.5 +/- 0.5
nmol/mg dry wt, respectively (P les than 0.001). Magnesium content of both
preparations was 37 nmol/mg dry wt. Potassium contents were 92 and 154
meq/l, respectively (P less than 0.001). Potassium content of
calcium-deficient cells increased to 161 meq/l following incubation for 30
min in a medium containing 1.6 mM ionized calcium. When incubated in a
medium containing a subphysiologic concentration of ionized calcium,
calcium-deficient cells rapidly lost the ability to exclude trypan blue and
to retain lactate dehydrogenase activity. As contrasted to
calcium-sufficient hepatocytes, calcium-deficient cells failed to
accumulate alpha-aminoisobutyric acid by active transport and lacked
microvilli and nuclear contents. This study supports simultaneous addition
of calcium and collagenase to the isolation medium as a means for
preserving physical, functional, and morphological integrity of isolated
hepatic parenchymal cells.
