Conditions affecting primary cell cultures of functional adult rat hepatocytes
- 51 Downloads
Primary monolayer cell cultures of adult rat hepatocytes underwent change in morphology and substantial cell loss between 1 and 3 days postinoculation. Dexamethasone-supplementation (1µM) of the culture medium maintained the polygonal epithelial morphology of the hepatocytes and increased longevity such that over 80% of the cells survived for 3 days and at least 30% for 8 or 9 days. This enhancement of survival was obtained up to 48 hr postinoculation, but the earlier the time of dexamethasone supplementation the greater the effect. Removal of dexamethasone resulted in a decrease in longevity. The positive effect of dexamethasone on longevity was observed following dexamethasone replacement of insulin in supplemented cultures, but the combination of insulin and dexamethasone resulted in poorer survival than with dexamethasone alone. The results are interpreted to indicate that dexamethasone provided a requirement of the in vitro environment for survival and suggest that elaboration of a complex medium is required to maintain hepatocytes in culture.
Key wordshepatocyte cultures dexamethasone insulin tissue culture liver perfusion
Unable to display preview. Download preview PDF.
- 1.Sandstrom, B. 1964. A method of cultivating morphologically highly differentiated liver epithelium in vitro. Acta Soc. Med. Upsal. 69:209–214.Google Scholar
- 6.Katsuta, H., and T. Takaoka. 1963. Carcinogenesis in tissue culture. II. Cultivation of normal rat liver cells. Japan J. Exp. Med. 33:265–275.Google Scholar
- 9.Coon, H. 1969. Clonal culture of differentiated cells from mammals: rat liver cell culture. Carnegie Inst. Washington Year Book 67:419–421.Google Scholar
- 19.Masuji, H., H. Nakabayashi, and J. Sato. 1974. Long-term cultivation of two diploid epithelial cell lines derived from normal rat liver cells. Acta Med. (Okayama) 28:281–293.Google Scholar
- 33.Billat, C., C. Plas, and R. Jacquot. 1972. Etude en microscopie electronique d’hepatocytes foetaux de rat en culture primaire: action du cortisol. Compt. Rend. 274:2338–2340.Google Scholar
- 35.Berliner, J. A. 1975. Effect of glucocorticoids on the ultrastructure of cultured liver cells. In: L. E. Gerschenson, and E. B. Thompson (Eds.),Gene Expression and Carcinogenesis in Cultured Liver. Academic Press, New York, pp. 62–93.Google Scholar
- 36.Gelehrter, T. D., W. L. Risser, and S. B. Reichberg. 1975. Hormonal regulation of amino acid transport in rat hepatoma cells in tissue culture. In: L. E. Gerschenson, and E. B. Thompson (Eds.),Gene Expression and Carcinogenesis in Cultured Liver. Academic Press, New York, pp. 190–204.Google Scholar
- 38.Harper, H. A. 1973.Review of Physiological Chemistry. Fourteenth edition. Lange Medical Publications, Los Altos, Calif.Google Scholar