. . "CZ - \u010Cesk\u00E1 republika" . "Optimizace kryoprezervace prase\u010D\u00EDch hepatocyt\u016F srovn\u00E1n\u00EDm \u017Eivotnosti a enzymatick\u00E9 aktivity \u010Derstv\u00FDch a kryoprezervovan\u00FDch bun\u011Bk"@cs . "Freezing conditions; MTT test; cytochrom P450; primary cultures; porcine hepatocytes"@en . . "Dvo\u0159\u00E1k, Zden\u011Bk" . . . "P(GA311/98/0648), Z(MSM 141100003), Z(MSM 151100003), Z(MZE-M03-99-01)" . . . . . "8"^^ . "Cryopreservation of porcine hepatocytes would ensure the accessibility of cells for laboratory use, permit the standardisation of experiments and save animals? lives. Therefore, in this study, we sought the optimal procedure for cryopreservation of porcine hepatocytes for both laboratory and clinical purposes. Hepatocytes were isolated from the liver lobe of a mini-pig by two-step collagenase perfusion. The cells were frozen with 20% foetal calf serum and 15% DMSO in two different media in four different concentrations ranging from 1 x 106 cells/ml to 5 x 106 cells/ml. 1.8 ml cryotubes and 120 ml Baxter bags were used for this purpose. Cells were cryopreserved either in a controlled freezer Sylab or step by step in a styrofoam box and stored at -196 \u00BAC. The quality of fresh and cryopreserved hepatocytes were assessed by trypan blue exclusion test and by the evaluation of cytochrome P450 isoenzymes and glutathione-S-transferase activities; primary cultures were evaluated morphologically and by"@en . "D\u00EDt\u011B, Petr" . . . "Optimisation of porcine hepatocyte cryopreservation by comparison of viability and enzymatic activity of fresh and cryopreserved cells" . "70" . "690305" . . "RIV/61989592:15110/01:00006958" . "7"^^ . "Acta Veterinaria Brno" . "Optimisation of porcine hepatocyte cryopreservation by comparison of viability and enzymatic activity of fresh and cryopreserved cells"@en . "3"^^ . "Optimisation of porcine hepatocyte cryopreservation by comparison of viability and enzymatic activity of fresh and cryopreserved cells"@en . . "Cryopreservation of porcine hepatocytes would ensure the accessibility of cells for laboratory use, permit the standardisation of experiments and save animals? lives. Therefore, in this study, we sought the optimal procedure for cryopreservation of porcine hepatocytes for both laboratory and clinical purposes. Hepatocytes were isolated from the liver lobe of a mini-pig by two-step collagenase perfusion. The cells were frozen with 20% foetal calf serum and 15% DMSO in two different media in four different concentrations ranging from 1 x 106 cells/ml to 5 x 106 cells/ml. 1.8 ml cryotubes and 120 ml Baxter bags were used for this purpose. Cells were cryopreserved either in a controlled freezer Sylab or step by step in a styrofoam box and stored at -196 \u00BAC. The quality of fresh and cryopreserved hepatocytes were assessed by trypan blue exclusion test and by the evaluation of cytochrome P450 isoenzymes and glutathione-S-transferase activities; primary cultures were evaluated morphologically and by" . "15110" . . . "RIV/61989592:15110/01:00006958!RIV09-MSM-15110___" . "Lata, Jan" . . . "Ulrichov\u00E1, Jitka" . . . "Smr\u017Eov\u00E1, Jana" . . "Machala, Miroslav" . "Optimizace kryoprezervace prase\u010D\u00EDch hepatocyt\u016F srovn\u00E1n\u00EDm \u017Eivotnosti a enzymatick\u00E9 aktivity \u010Derstv\u00FDch a kryoprezervovan\u00FDch bun\u011Bk"@cs . "[429A36C4CC26]" . "0001-7213" . "Bl\u00E1ha, Lud\u011Bk" . "\u0160im\u00E1nek, Vil\u00EDm" . "2" . . "Optimisation of porcine hepatocyte cryopreservation by comparison of viability and enzymatic activity of fresh and cryopreserved cells" . . "Kryoprezervace prase\u010D\u00EDch hepatocyt\u016F by zabezpe\u010Dila dostupnost bun\u011Bk pro laboratorn\u00ED u\u017Eit\u00ED, dovolila standardizaci experiment\u016F a u\u0161et\u0159ila \u017Eivoty zv\u00ED\u0159at. V t\u00E9to pr\u00E1ci jsme proto hledali nejlep\u0161\u00ED postup kryoprezervace prase\u010D\u00EDch hepatocyt\u016F jak pro laboratorn\u00ED, tak pro klinick\u00E9 \u00FA\u010Dely. Hepatocyty byly izolov\u00E1ny z jatern\u00EDho laloku experiment\u00E1ln\u00EDho miniaturn\u00EDho prasete pomoc\u00ED dvoustup\u0148ov\u00E9 perf\u00FAze kolagen\u00E1zou. Bu\u0148ky byly pot\u00E9 zamra\u017Eeny ve 20% fet\u00E1ln\u00EDm telec\u00EDm s\u00E9ru a 15% DMSO ve 2 r\u016Fzn\u00FDch m\u00E9di\u00EDch ve 4 koncentrac\u00EDch od 1 x 106 bun\u011Bk/ml do 5 x 106 bun\u011Bk/ml. Pou\u017Eity byly kryotuby o objemu 1.8 ml a vaky Baxter o objemu 120 ml. Bu\u0148ky byly zamra\u017Eeny bu\u010F \u0159\u00EDzen\u00FDm mra\u017Een\u00EDm v p\u0159\u00EDstroji Sylab, nebo postupn\u00FDm ochlazov\u00E1n\u00EDm v polystyr\u00E9nov\u00FDch boxech a pot\u00E9 uchov\u00E1v\u00E1ny p\u0159i -196 \u00BAC. Kvalita \u010Derstv\u00FDch a kryoprezervovan\u00FDch hepatocyt\u016F byla hodnocena v testu s trypanovou mod\u0159\u00ED a m\u011B\u0159en\u00EDm aktivity izoenzym\u016F cytochromu P450 a glutathion-S-transfer\u00E1zy; prim\u00E1rn\u00ED kultury byly posuzov\u00E1ny morfologicky a testem MTT. Kryoprezervovan\u00E9 h"@cs . .