Digitonin-collagenase perfusion for efficient separation of periportal or perivenous hepatocytes.
نویسندگان
چکیده
Intact rat liver cells from the perivenous region were isolated by collagenase perfusion after first destroying the periportal region by a brief portal infusion of digitonin. Periportal cells were isolated after retrograde digitonin infusion. Significantly higher alanine aminotransferase, gamma-glutamyltransferase and lactate dehydrogenase activities and lower glutamate dehydrogenase and pyruvate kinase activities in periportal than in perivenous cells demonstrate marked separation. The high yield allows further characterization in vitro of the cell populations.
منابع مشابه
Urea synthesis in freshly isolated and in cultured periportal and perivenous hepatocytes.
Periportal hepatocytes isolated by digitonin/collagenase perfusion produced urea faster than did similarly prepared perivenous hepatocytes, in both the presence and the absence of amino acids and various urea precursors. There was no difference between the two cell types in rates of intracellular proteolysis. The initial difference in urea synthesis persisted for 5 days during primary culture, ...
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Hepatocytes in the afferent (periportal) and efferent (perivenous) acinar zones have different morphological and biochemical characteristics [ I ] . Periportal hepatocytes have a greater capacity for gluconeogenesis and urea synthesis, and a lower capacity for lipogenesis and glutamine formation compared to perivenous hepatocytes [I]. The pathways of detoxification such as monooxygenation and s...
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The zonal distribution of GSH metabolism was investigated by comparing hepatocytes obtained from the periportal (zone 1) or perivenous (zone 3) region by digitonin/collagenase perfusion. Freshly isolated periportal and perivenous cells had similar viability (dye exclusion, lactate dehydrogenase leakage and ATP content) and GSH content (2.4 and 2.7 mumol/g respectively). During incubation, perip...
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The mammalian liver microcirculation seems to be uniquely organized in order to create maximum difference of the biochemical milieu to which the upstream (periportal) and downstream (perivenous) parenchymal cells are exposed. Indeed the existence of a subdivision of hepatocytes in zones of different metabolic function, long predicted by enzyme zonation studies (for a review, see Jungermann & Ka...
متن کاملDistribution of albumin, alpha 1-inhibitor 3 and their respective mRNAs in periportal and perivenous rat hepatocytes isolated by the digitonin-collagenase technique.
The expression of albumin and alpha 1-inhibitor 3 genes was investigated in rat cell suspensions enriched in periportal (n = 10) and perivenous (n = 10) hepatocytes obtained by the digitonin-collagenase technique. The degree of enrichment of the cell suspensions was assessed: (1) by enzymic assays for the periportal marker alanine aminotransferase and for the perivenous marker glutamine synthet...
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عنوان ژورنال:
- The Biochemical journal
دوره 228 3 شماره
صفحات -
تاریخ انتشار 1985