Preparation of rat liver cells. III. Enzymatic requirements for tissue dispersion

摘要: Abstract Isolated perfused rat livers were dispersed by a two-step procedure of Ca2+ removal (probably including the Ca2+-dependent adhesion factor) followed enzymatic treatment. Collagenase, crude or purified, converted all parenchymal tissue to suspension cells which 95% intact. Other enzymes such as hyaluronidase, lysozyme and trypsin ineffective. Since hypoxic conditions during treatment did not affect cellular viability, oxygenation apparatus can be omitted facilitate sterile preparation cells. Purification differential centrifugation reduced contamination non-parenchymal from 10–20% initially 1–2% finally, with 40% recovered no loss in viability. The purified 20% binucleated had same biochemical composition intact liver tissue. active synthesis RNA, protein, glycogen various metabolites, showed sensitivity towards steroid polypeptide hormones. Many freshly isolated marked constriction furrows disappeared upon incubation.