A method of obtaining a suspension of intact parenchymal cells from adult rat liver

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Abstract
The perfusion of liver with either citrate or tetraphenyl boron to remove Ca2+ or K+ or with a solutionof high osmolarity and alkaline pH yields plenty of cells but they are all damaged.Perfusion of the liver with hyaluronidase and collagenase followed by incubation of liver slices in the same enzyme solution produced a high yield of cells (25 %, w/w, of liver) of which only about 1 % were undamaged. However, perfusion with 0·3 % hyaluronidase, 0·3 % collagenase and 0·1 % trypsin in phosphate-buffered saline (excluding Mg2+ and Ca2+ followed by incuba-tion at 25 °C of the chopped liver gave a small yield (2–4%, w/w) of undamaged cells which were not permeable to eosin for up to an hour when suspended in culture medium containing 2 % bovine serum albumin.