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Separation of adult rat hepatocytes into distinct subpopulations by centrifugal elutriation. Morphological, morphometrical and biochemical characterization of cell fractions

par Bernaert, Denise ;Wanson, Jean-Claude ;Mosselmans, Roger
Référence PASCAL. F 52, Biochimie, biophysique moléculaire, biologie moléculaire et cellulaire, 34, 2-3, page (159-174)
Publication Publié, 1979

Article révisé par les pairs

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Résumé :

Isolation hepatocytes from adult rats were separated by elutriation (counterflow centrifugation) into various intact subpopulations of different cell sizes. The morphological, morphometrical and cytophotometrical characteristics of the elutriated hepatocytic subpopulations were determined. Hepatocytes recovered in the different classes display a well preserved ultrastructure. Their degree of ploidy is related to their cell size. Their functional integrity is attested to by high levels of glucose-6-phosphatase and glycogen synthetase activities and by the capacity to stimulate the active form of synthetase, in the presence of glucose. Our results show that the elutriation process is able to separate: Most of the cell debris and damaged hepatocytes from the intact ones; Well preserved isolated hepatocytes of different diameters and degree of ploidy: 17.5 μm (2 n); 19.9 μm and 20.8 μm (4 n); 24.6 μm (8 n); and perfectly isolated cells from doublets, triplets or aggregates.