The viability of parenchymal liver cells isolated from young and old rats
article
Liver parenchymal cells were isolated from female WAG/Rij rats, aged from 2 wk to 36 mth, by means of collagenase and hyaluronidase. The isolated cells met the following morphological and biochemical criteria for viability. The impermeability of the plasma membrane was demonstrated by trypan blue exclusion. A high percentage of the cells remained impermeable to trypan blue during storage at 4°C; however, under experimental conditions at 37°C, the percentage of impermeable cells declined. The ultrastructural integrity of the impermeable cells was confirmed by electron microscopic observations. As a test for the metabolic integrity of the cells, measurements were performed on their endogenous respiration. The oxygen consumption was comparable to that of hepatic parenchymal cells in intact tissue. Succinate or other tricarboxylic acid cycle intermediates did not increase the respiration of isolated cells, but a marked stimulation was observed if cell debris was present. The endogenous respiration per million cells was higher for cells from old rats than for cells from young rats. This increase could be attributed to an increase in average cell size resulting from a shift to polyploid cells with age. The availability of viable liver cells from both young and old rats offers the possibility of performing quantitative aging research on a cellular level.
Chemicals/CAS: hyaluronidase, 9001-54-1, 9055-18-9; Culture Media; Proteins; Succinates; Trypan Blue, 72-57-1
Chemicals/CAS: hyaluronidase, 9001-54-1, 9055-18-9; Culture Media; Proteins; Succinates; Trypan Blue, 72-57-1
Topics
clostridiopeptidase ahyaluronidaseagecellcell membranehistologyisolationliver cellparenchymarattheoretical studyAgingAnimalCell Membrane PermeabilityCells, CulturedChromosomesCulture MediaDiploidyFemaleLiverMicroscopy, ElectronOxygen ConsumptionPolyploidyProteinsRatsRats, Inbred StrainsStaining and LabelingSuccinatesTime FactorsTrypan Blue
TNO Identifier
227729
ISSN
00476374
Source
Mechanisms of Ageing and Development, 3(2), pp. 107-119.
Pages
107-119
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