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Journal of Cell Science, Vol 8, 73-86, Copyright © 1971 by Company of Biologists

Submitted on May 4, 1970

A Method of Obtaining a Suspension of Intact Parenchymal Cells from Adult Rat Liver

JENNIFER J. GALLAI-HATCHARD 1 and G. M. GRAY 1

1 Lister Institute of Preventive Medicine, Chelsea Bridge Road, London, S.W.1, England

The perfusion of liver with either citrate or tetraphenyl boron to remove Ca2+ or K+ or with a solution of 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 incubation 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.

Submitted on May 4, 1970






© The Company of Biologists Ltd 1971