Experiments on an ascites hepatoma: I. Enzymatic digestion and alkaline degradation of the cementing substance and separation of cells, in tumor islands

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Summary o 1. Various types of hydrolytic enzymes were tested for ability to digest the intercellular cementing substance in islands of two ascites hepatomas. Three preparations of hyaluronidase, as well as other carbohydrases and a lipase, failed to cause separation of the constituent cells. 2. Preparations of crude trypsin, crystalline trypsin and crystallinechymotrypsin, in phosphate buffer at pH 7.3 to 8.0, effectively separated cells in many hepatoma islands. This reaction was usually characterized by the formation of a mucous-like coagulum containing almost all of the freed cells. Similarly, Polidase-S, animal diastase, and alpha amylase were effective at pH 7.3 to 8.0. 3. The action of trypsin was due in part to the presence of phosphate. Thus, incubation of tumor islands in phosphate buffer without enzyme, caused a loosening and partial separation of tumor cells. 4. Various enzyme-free salt solutions possessing no action against the intercellular matrix at pH 7.0 to pH 9.0, were effective when the pH was above 9.0. The separation of tumor cells in alkaline solution was equal to, and often more complete, than in the case of trypsin. 5. These data suggest that the ability of the intercellular cementingsubstance in maintaining the integrity of the hepatoma island may depend largely upon the presence of protein.