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- Belfrage, Per, et al.
(författare)
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Dispersion of viable pig liver cells with collagenase
- 1975
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Ingår i: Life Sciences. - : Elsevier BV. - 1879-0631 .- 0024-3205. ; 17:8, s. 1219-1225
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Tidskriftsartikel (refereegranskat)abstract
- Viable suspended hepatocytes were prepared from surgical biopsy specimens of pig and human liver by digestion with collagenase. Initial perfusion of the tissue through cannulated blood vessels with 0.5 mM EGTA followed by 0.2% collagenase gave the best results. 20−870 × 106 cells of which 60–95 % excluded trypan blue were obtained from 5–30 g pig liver pieces, while results with human liver specimens were usually less satisfactory. In some experiments, however, viable cells, as judged by vital stain exclusion and ability to synthesize lipids were obtained in sufficient yield. In the pig hepatocytes glycerolipid synthesis from [3H] glycerol and oxidation and esterification of [14C] oleic acid had the same characteristics as those observed earlier in rat hepatocytes.
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- Florén, Claes-Henrik, et al.
(författare)
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Binding, interiorization and degradation of cholesteryl ester labelled chylomicron remnant particles by rat hepatocyte monolayers
- 1977
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Ingår i: The Biochemical journal. - : Portland Press Ltd.. - 0264-6021. ; 168:3, s. 483-494
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Tidskriftsartikel (refereegranskat)abstract
- 1. The cholesteryl ester of isolated chylomicron-remnant particles was efficiently degraded by hepatocyte monolayers. The degradation was sensitive to metabolic inhibitors. 2. With increasing amounts of remnant cholesteryl ester the rate of uptake approached saturation and conformed to a linear double-reciprocal plot. The V(max.) was determined as 80ng of cholesteryl ester/h per mg of protein and the apparent K(m) as 1.4mug of cholesteryl ester per mg of protein. The time course for the uptake and hydrolysis suggested that binding of particles to the cell surface preceded the degradation. 3. Cholesteryl esters of native chylomicrons were degraded to a much smaller extent and their presence had only a small inhibitory effect on the degradation of chylomicron remnants. Intestinal very-low-density lipoproteins were degraded somewhat faster than chylomicrons, and caused more inhibition of remnant degradation. Rat high-density lipoproteins inhibited the hydrolysis of remnant cholesteryl ester by up to 50%, but had less influence on the amount of cholesteryl ester that was bound to the cells. Serum decreased both the uptake and hydrolysis, whereas d=1.21 infranatant had no effect. 4. The cholesteryl ester hydrolysis after the uptake by the cells was inhibited by chloroquine and by colchicine. Only 28-36% of the unhydrolysed cholesteryl ester could be released from these cells by trypsin treatment, indicating that the major portion was truly intracellular. The particles that could be released from the cell surface by trypsin and those remaining in the medium had the same triacylglycerol/cholesteryl ester ratio as the added remnant particles. Significant amounts of denser particles were thus not formed during contact with the cell surface. 5. The presence of heparin, as well as preincubation of the cells with heparin, increased the uptake of chylomicron remnants. This effect was most marked in the presence of serum. A much smaller proportion of the other serum lipoproteins was taken up, and this proportion was not increased by heparin.
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- Florén, Claes-Henrik, et al.
(författare)
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Effects of fatty acid unsaturation on chylomicron metabolism in normal and hepatectomized rats
- 1977
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Ingår i: Eur J Biochem. - : Wiley. - 0014-2956. ; 77:1, s. 23-30
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Tidskriftsartikel (refereegranskat)abstract
- 1. Hepatectomized rats were injected intravenously with doubly labelled ([14C]linoleic acid and [3H]palmitic acid) thoracic duct lymphs from rats fed cream, triolein or corn oil. The disappearance of the radioactive fatty acids of different molecular triacylglycerol species and of phospholipids from plasma was studied.2. 73–93% of the injected triacylglycerols had been cleared from plasma within 15 min. At all stages of lipolysis the 3H/14C ratio of the plasma triacylglycerol was the same as in the injected material. If the cream chyle had been cooled to 4 °C before use there was, however, an enrichment of [3H]palmitic acid and of fully saturated triacylglycerols in the remnant particles formed.3. Only 38–50% of the radioactive chyle phosphatidylcholine was eliminated from plasma in 30 min. At this time most of the remaining phosphatidylcholine was, however, in other lipo‐protein classes than the chylomicron remnants.4. Also in intact rats data were obtained, indicating that the major portion of chylomicron phospholipids is transferred to other serum lipoproteins by exchange or net movement rather than being hydrolysed in the 1‐position by lipoprotein lipase or taken up intact by the liver.5. More of both the labelled fatty acids appeared in liver triacylglycerols in experiments with cream chyle than in experiments with corn oil chyle. Data were obtained suggesting that this may be due to a higher uptake of intact triacylglycerol as remnant particles.6. When linoleic acid is fed as a tracer dose in cream, a high proportion (16–36%) is incorporated into chyle phospholipids.
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