A morphological study of cooling rate response in normal and neoplastic human liver tissue: cryosurgical implications

The process of freezing in normal human livers and in human liver tumors was studied by freezing samples of these tissues with constant cooling rates and then examining the morphology of the frozen tissue, after freeze substitution, with the light microscope. Cooling rates varied from 2 degrees C/min up to approximately 2000 degrees C/min. It was observed that high cooling rates produce extensive intracellular ice in both normal and neoplastic liver. At slow rates of cooling, normal and neoplastic liver cells dehydrated and large extracellular ice crystals formed. Comparison of the frozen normal liver and the frozen malignant tumors shows that for the same rates of freezing, the tumor cells retain more cellular water and therefore show less susceptibility to dehydration at low rates of cooling. At slow cooling rates, the amount of cellular dehydration and consequent vascular and interstitial space engorgement changed with the type of tissue frozen. The greatest amount of dehydration occurred in normal human liver, followed by metastatic colon carcinoma and finally primary hepatocellular carcinoma. These results are important for cryosurgery since they suggest that malignant tissues have a different response to freezing than normal tissues.