1988
DOI: 10.1016/0041-008x(88)90081-6
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In vitro toxicity assessment of cyclosporin a and its analogs in a primary rat hepatocyte culture model

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Cited by 25 publications
(6 citation statements)
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“…Isolation and Culture of Hepatocytes. Hepatocytes were isolated from rats anesthetized with sodium pentobarbital (Nembutal, 50 mgkg body weight, intraperitoneally; University Hospital, Zurich, Switzerland) with a modified two-step collagenase perfusion method (12) as described earlier (13). First, the liver was perfused in situ with a Ca2+-free, hemoglobin-free and phenol red-free Hanks' bicarbonate buffer (supplemented with 0.5 mmol/L EGTA and 20 mmol/L HEPES) for 5 min at 37" C. The liver was then removed, and the perfusion was continued with an oxygenated (95% 0,; 5% CO,) Hanks' buffer containing Ca2' (5 mmoVL), NaHCO, (25 mmol/L), BSA (0.05%) and collagenase (type I-CLD; 100 U/ml) for 10 min (flow rate = 30 ml/min).…”
Section: Chemicalsmentioning
confidence: 99%
“…Isolation and Culture of Hepatocytes. Hepatocytes were isolated from rats anesthetized with sodium pentobarbital (Nembutal, 50 mgkg body weight, intraperitoneally; University Hospital, Zurich, Switzerland) with a modified two-step collagenase perfusion method (12) as described earlier (13). First, the liver was perfused in situ with a Ca2+-free, hemoglobin-free and phenol red-free Hanks' bicarbonate buffer (supplemented with 0.5 mmol/L EGTA and 20 mmol/L HEPES) for 5 min at 37" C. The liver was then removed, and the perfusion was continued with an oxygenated (95% 0,; 5% CO,) Hanks' buffer containing Ca2' (5 mmoVL), NaHCO, (25 mmol/L), BSA (0.05%) and collagenase (type I-CLD; 100 U/ml) for 10 min (flow rate = 30 ml/min).…”
Section: Chemicalsmentioning
confidence: 99%
“…Livers were obtained from 150 to 200 g male Sprague-Dawley rats and hepatocytes isolated by a standardized Ca z+ free collagenase perfusion technique as previously described (23) and modified (24). The cells (viability > 85% by trypan blue exclusion) were plated on collagen type I (Serva, Heidelberg, Germany) coated plastic dishes (Falcon Primaria, Becton Dickinson Labware, Oxnard, CA) at a density of 7 x 10 4 cells/cm 2 and cultured in phenol red free Williams E medium (Animed, Basel, Switzerland) supplemented with penicillin (100 U/ml), streptomycin (0.1 mg/ml), insulin (10~7 M), dexamethasone (10~7 M), and 10% fetal calf serum.…”
Section: Preparation Of Hepatocyte Culturesmentioning
confidence: 99%
“…However, such selective inhibitors are not yet available for many of the transporters relevant in drug distribution and elimination (Hirano et al, 2006;Webborn et al, 2007). In addition they might also influence many other processes in the cell systems used (Boelsterli et al, 1988;Ratanasavanh et al, 1996). Optionally, passive processes can be assessed by including in the experiment high concentrations in which active transport processes are saturated, allowing the determination of passive diffusion processes Yamashiro et al, 2006).…”
mentioning
confidence: 99%