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Dispersion of viable pig liver cells with collagenase.
Authors:Per Belfrage  Bengt Börjesson  Inga Hägerstrand  Åke Nilsson  Arne Olsson  Thomas Wiebe  Björn Åkesson
Institution:1. Department of Physiological Chemistry, University of Lund, Lund, Sweden;2. Department of Surgery, Hospital of Lund, University of Lund, Lund, Sweden;3. Institute of Pathology, Malmö General Hospital, University of Lund, Lund, Sweden
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.
Keywords:To whom reprint requests should be addressed  
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