Preparation of single cell suspension from rat liver. A simple method designed for immunologic studies.

Single cell suspensions from rat liver were prepared by mean of a 10 min. in vivo liver perfusion with an isotonic solution. This solution was devoid of calcium but contained 27 mM of sodium citrate and 1% of bovine serum albumin. The liver cells were dissociated over chromium nickel screen in presence of 0.8 mM of calcium and 0.25 M of saccharose; they were further separated by gravity sedimentation for 10 min. The dissociated cells, when incubated in Waymouth's medium enriched with 15% of heat-inactivated horse serum and 1% of BSA, actively synthetized urea and proteins. They also incorporated [3H] thymidine. Isolated hepatocytes, obtained without resorting to extraneous enzymes, conform to the conditions required for the study of cell surface receptors involved in immunologic recognition mechanisms.