Spontaneous51Cr release by isolated rat hepatocytes: An indicator of membrane damage

Zawydiwski, R.; Duncan, Robin E.

doi:10.1007/bf02616167

Cited by 37 publications

(10 citation statements)

References 28 publications

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“…In the presently described model as many as 20% of the injected hepatocytes are retained by the recipient liver in The 51Cr label used in our experiments functions as a cellular marker that is rapidly lost and not reutilised upon cellular lysis (Zawydiwski & Duncan, 1978); retention of 51Cr therefore reflects the retention of intact cells. The stability of the marker is illustrated by our results (Figure 1).…”

Section: Discussionmentioning

confidence: 97%

Diploid nature of hepatocellular tumours developing from transplanted preneoplastic liver cells

Sæter

Schwarze²,

Nesland³

et al. 1989

Br J Cancer

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Pitot, 1985;Saeter et al., 1987;Roomi et al., 1985;Hunt et al., 1982). Transplantation experiments permit studies of individual separable cell subpopulations generated during carcinogenesis as well as of the behaviour of carcinogen-altered cells in an in vivo environment not exposed to carcinogens (Hanigan & Pitot, 1985).One interesting feature of carcinogen-altered hepatocytes is their change in DNA content. We have previously reported a significant increase in the fraction of diploid hepatocytes during early stages of liver carcinogenesis induced by treatment with diethylnitrosamine (DEN) and 2-acetylaminofluorene (AAF) (Schwarze et al., 1984;Seglen et al., 1988b). Moreover, neoplastic nodules and hepatocellular carcinomas generated in this model have been shown to contain 70-90% diploid cells, as compared to only 10% in the normal, polyploid liver Saeter et al., 1988a). Similar findings have been reported in other models of rat liver carcinogenesis (Neal et al., 1976; Irving et al., 1977;Styles et al., 1985;Deleener et al., 1987), indicating that replacement of normal polyploidising growth by diploid divisional proliferation may be a fundamental feature of chemical hepatocarcinogenesis.To further investigate the stability and importance of this phenotypic alteration we have studied the DNA content of isolated nuclei from neoplastic nodules and hepatocellular carcinomas arising in host liver after intraportal injection of hepatocytes from syngeneic carcinogen-treated donor rats. Finally, we have characterised our transplantation model in terms of degree of donor cell retention in host liver and cell dose versus tumour yield, and studied the effect of secondary promotion in the host with dietary phenobarbitone (PB). Materials and methodsDonor animal treatment and isolation of donor hepatocytes Four-week old male rats (70g) of the inbred Wistar Kyoto strain were subjected to partial hepatectomy (PH) and 24h later injected intraperitoneally with DEN (50mgkg-1). Following one week's rest on basal diet, the animals were fed a semi-synthetic diet (Bio-Serv Inc., Frenchtown, NJ, USA) containing 0.02% AAF for 4 weeks, then returned to basal diet. This initiation-promotion regimen produces multiple neoplastic nodules from 8 weeks after start of treatment and hepatocellular carcinomas from 4 months onwards . At 6 or 8 weeks after start of treatment donor hepatocytes were isolated by two-step collagenase liver perfusion and the cells purified by differential centrifugation as described previously (Seglen, 1976). Some donor cell suspensions from carcinogen-treated animals were subjected to centrifugal elutriation for the purpose of altering the relative amounts of diploid and polyploid donor hepatocytes before injection into the host liver. The viability of donor hepatocytes was in general in excess of 90% as determined by trypan blue exclusion.For control experiments and studies of donor cell retention in recipient liver, donor hepatocytes were obtained by collagenase perfusion of livers from normal, untreated 10-week...

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Section: Discussionmentioning

confidence: 97%

Diploid nature of hepatocellular tumours developing from transplanted preneoplastic liver cells

Sæter

Schwarze²,

Nesland³

et al. 1989

Br J Cancer

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“…The effect of LfcinB on HUVEC viability was assessed by 51 Cr release from the intracellular compartment 32 and Hoechst 33342 trihydrochloride dye staining of nuclear material. 33 For 51 Cr release, HUVECs were labeled for 1 hour with 100 Ci of Na 2 51 CrO 4 (MP Biomedicals).…”

Section: Cell Viability Assaysmentioning

confidence: 99%

Bovine Lactoferricin Inhibits Basic Fibroblast Growth Factor- and Vascular Endothelial Growth Factor165-Induced Angiogenesis by Competing for Heparin-Like Binding Sites on Endothelial Cells

Mader

Smyth

Marshall

et al. 2006

The American Journal of Pathology

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“…The cultures were incubated for 4 h at 37°C. First, 50 tzl of the supernatant without cells were sampled to check the viability of the lymphocytes [27]. This cell viability test relies on the fact that chromate is taken up by living cells and reduced by the metabolism.…”

Section: Lymphocyte Adhesion Assaymentioning

confidence: 99%

A ground‐based model to study the effects of weightlessness on lymphocytes

Gmünder

Kiess

Sonnefeld

et al. 1990

Biology of the Cell

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The mitogenic response of human lymphocytes was found to be markedly reduced in weightlessness conditions as compared to normal gravity. One possible explanation is that due to the non-existent sedimentation in space the lymphocytes could not adhere and spread on a substratum. Thus, we investigated the effect of substratum adhesiveness on lymphocyte responsiveness by reducing and blocking cell adhesion with poly-HEMA in a simple on-ground system. Lymphocyte adhesiveness was assessed by measuring the proportion of non-adhesive, slightly, and strongly adhesive 51Cr-radiolabelled cells on uncoated and poly-HEMA coated plastic. The amount of cell spreading on surfaces with varying adhesiveness was determined by measuring the area of cells. Cells grown on medium and thick poly-HEMA films were rounded in shape. By contrast, on tissue culture plastic, they showed clear signs of spreading. The mitogenic response of lymphocytes grown on thick poly-HEMA films was reduced by up to 68% of the control (tissue culture plastic). Interferon-gamma production was virtually nil when the cells were grown on the least adhesive substratum. These results show that activated lymphocytes need to anchor and spread prior to achieving an optimal proliferation response. We conclude that decreased lymphocyte adhesion could contribute to the depressed in vitro lymphocyte responsiveness found in the microgravity conditions of space flight.

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Spontaneous51Cr release by isolated rat hepatocytes: An indicator of membrane damage

Cited by 37 publications

References 28 publications

Diploid nature of hepatocellular tumours developing from transplanted preneoplastic liver cells

Diploid nature of hepatocellular tumours developing from transplanted preneoplastic liver cells

Bovine Lactoferricin Inhibits Basic Fibroblast Growth Factor- and Vascular Endothelial Growth Factor165-Induced Angiogenesis by Competing for Heparin-Like Binding Sites on Endothelial Cells

A ground‐based model to study the effects of weightlessness on lymphocytes

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