Ultrastructural study of isolated rat hepatocyte suspensions incubated in incomplete or complete culture medium

Ultrastructural changes in isolated rat hepatocytes (IHC) in suspension were examined after 1, 3, 5 and 10 hr incubation, and the influence of Krebs-Henseleit (K-H) buffer as an incomplete medium and GIT culture medium (Nihon Pharmaceutical Co.) as a complete culture medium were investigated. The viability was approximately 60-75% after 1, 3 and 5 hr incubation, and was slightly higher in GIT medium than K-H buffer. Immediately after isolation, IHC showed a cuboidal shape, and after 3 and 5 hr incubation, a round shape with numerous surface microvilli. Cytoplasmic organelles appeared almost normal. After 10 hr incubation, cells reaggregated and the number of autophagic vacuoles increased. Cellular changes, such as slight dilatation of the endosplasmic reticulum and Golgi apparatus, and bleb formation in the cell surface, were more often observed in IHC incubated in K-H buffer than in GIT medium. IHC incubated in GIT medium contained glycogen particles in the cytoplasm, and lipoprotein-like particles were observed in the cisternae of Golgi apparatus and smooth endoplasmic reticulum. The lipoprotein-like particles were found in the intercellular spaces after 10 hr incubation. These results suggest that GIT medium is superior to K-H buffer in its ability to preserve normal structure and function of IHC.