Summary
Freshly harvested rat hepatocytes form spheroids on uncoated positively charged polystyrene surfaces. Time lapse microscopy revealed that cell movement and reorganization were involved in spheroid formation. Ultrastructural evaluation using scanning and transmission electron microscopy indicated polarized cellular morphology and extensive cell-cell communication within spheroids. Bile canalicular structures were observed to surround each individual hepatocyte, forming an intricate three-dimensional continuous network of channels that appeared to end as pores/holes on the surface of the spheroid. The maintenance of differentiated cellular morphology coincided with preservation of hepatocyte viability and enhanced levels of tissue specific functions in spheroids.
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Asano, K.; Koide, N.; Tsuji, T. Ultrastructure of multicellular spheroids formed in the primary culture of adult rat hepatocytes. J. Clin. Electron Microsc. 22:243–252; 1989.
Barcellos-Hoff, M. H.; Aggeler, J.; Ram, T. G., et al. Functional differentiation and alveolar morphogenesis of primary mammary cultures on reconstituted basement membrane. Development 105:223–235; 1989.
Ben-Ze’ev, A.; Robinson, G. S.; Bucher, N. L., et al. Cell-cell and cell-matrix interactions differentially regulate the expression of hepatic and cytoskeletal genes in primary cultures of rat hepatocytes. Proc. Natl. Acad. Sci. USA 85:2161–2165; 1988.
Halban, P. A.; Powers, S. L.; George, K. L., et al. Spontaneous reassociation of dispersed adult rat pancreatic islet cells into aggregates with three-dimensional architecture typical of native islets. Diabetes 36:783–790; 1987.
Koide, N.; Asano, K.; Sakaguchi, K., et al. Electron microscopic observation of vermipodia-like processes of multicellular spheroids formed in the primary culture. J. Clin. Electron Microsc. 22:5–6; 1989.
Koide, N.; Sakaguchi, K.; Koide, Y., et al. Formation of multicellular spheroids composed of adult rat hepatocytes in dishes with positively charged surfaces and under other nonadherent environments. Exp. Cell Res. 186:227–235; 1990.
Koide, N.; Shinji, T.; Tanabe, T., et al. Continued high albumin production by multicellular spheroids of adult rat hepatocytes formed in the presence of liver-derived proteoglycans. Biochem. Biophys. Res. Commun. 161:385–391; 1989.
Nyberg, S. L.; Shatford, R. A.; Peshwa, M. V., et al. Evaluation of hepatocyte function in a bioartificial liver: a potential device for the treatment of liver failure. Biotechnol. Bioeng. 41:194–203; 1993.
Nyberg, S. L.; Shirabe, K.; Peshwa, M., et al. Extracorporeal application of a gel-entrapment, bioartificial liver: demonstration of drug metabolism and other biochemical functions. Cell Trnasplantation 2:441–452; 1993.
Peshwa, M. V.; Wu, F. J.; Follstad, B. D., et al. Kinetics of hepatocyte spheroid formation. Biotechnol. Prog. 10:460–466; 1994.
Seglen, P. O. Preparation of isolated rat liver cells. Methods Cell Biol. 13:29–38; 1976.
Shannon, J. M.; Mason, R. J.; Jennings, S. D. Functional differentiation of alveolar type II epithelial cells in vitro: effects of cell shape, cell-matrix interactions and cell-cell interactions. Biochim. Biophys. Acta 931:143–156; 1987.
Shinji, T.; Koide, N.; Tsuji, T. Glycosaminoglycans partially substitute for proteoglycans in spheroid formation of adult rat hepatocytes in primary culture. Cell Struct. Funct. 13:179–188; 1988.
Tong, J. Z.; De, L. P.; Furlan, V., et al. Long-term culture of adult rat hepatocyte spheroids. Exp. Cell Res. 200:326–332; 1992.
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Peshwa, M.V., Wu, F.J., Sharp, H.L. et al. Mechanistics of formation and ultrastructural evaluation of hepatocyte spheroids. In Vitro Cell.Dev.Biol.-Animal 32, 197–203 (1996). https://doi.org/10.1007/BF02722946
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DOI: https://doi.org/10.1007/BF02722946