Abstract
Primary cultured rat hepatocytes in a membrane-supported collagen sandwich maintained their normal cell morphology and high level of albumin secretion for over 56 days. It was found that the existence of an upper layer of collagen gel is crucial for long-term culture and that the transference of cellular nutrients between the culture media and hepatocytes from both the upper and the lower sides of gel layers promotes albumin secretion. These facts suggest that the membrane-supported collagen sandwich mimics well thein vivo environment of hepatocytes. This method has great potential for the long-term culture of primary cells.
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References
Bissel DM, Arenson DM, Maher JJ and Roll FJ (1987) Support of cultured hepatocytes by a laminin-rich gel. Evidence for a functionally significant subendothelial matrix in normal rat liver. J. Clin. Invest. 79: 801–812.
Charmbard M, Gabrison J and Mauchamp J (1981) Influence of collagen gel on the orientation of epithelial cell polarity. J. Cell Biol. 91: 157–166.
Chen J, Stuckey EC, Berry CL (1985) Three-dimensional culture of rat exocrine pancreatic cells using collagen gels. Br. J. Exp. Pathol. 66: 551–559.
Dunn JCY, Tompkins RG, Yarmush ML (1991) Long-term in vitro function of adult hepatocytes in a collagen sandwich configuration, Biotechnology progress. 7: 237–245.
Dunn JCY, Yarmush ML, Koebe Hans G and Tompkins RG (1989) Hepatocyte function and extracellular matrix geometry: long-term culture in a sandwich configuration FASB J. 3: 174–177.
Guguen-Guillouzo C, Clement B, Baffet G, Beaumont C, Morel-Chany E, Glaise D and Guillouzo A (1983) Maintenance and reversibility of active albumin secretion by adult rat hepatoceytes co-cultured with another liver epithelial cell type. Exp. Cell Res. 143: 47–54.
Hamada S, & Fujita S (1983) DAPI staining improved for quantitative cytofluorometry. Histochemistry. 79: 219–226.
Ingber DE, Madri JA and Jamieson JD (1986) Basement membrane as a spatial organizer of polarized epithelia. Am. J. Pathol. 122: 129–139.
Martin GR (1987) Laminin and other basement membrane components. Annu. Rev. Cell. Biol. 3: 57–85.
Mesnil M, Fraslin JM, Piccoli C, Yamasaki H and Guguen-Guillouzo C (1987) Cell contact but not junctional communication (Dye Coupling) with biliary epithelial cells is required for hepatocytes to maintain. Exp. Cell Res. 173: 524–533.
Michalopoulos G and Pitot NC (1975) Primary culture of parenchymal liver cells on collagen membranes Exp. Cell Res. 94: 70–78.
Montesano R, Mouron P, Amherdt M and Orci L (1983) Collagen matrix promotes reorganization of pancreatic endocrine cell monolayers into islet-like organoids. J. Cell Biol. 97: 935–955.
Reid LM, Gatmaitan Z, Aris I, Ponce P and Rojkind M (1980) Long-term cultures of normal rat hepatocytes on liver biomatrix. Ann. N.Y. Acad. Sci. 349: 70–76.
Seglen PO (1976) Preparation of isolated rat liver cells. Methods Biol. 13: 29–83.
Sirca AE, Richards W, Tsukada Y, Satter CA and Pitot HC (1979) Fetal phenotypic expression by adult rat hepatocytes on collagen gel/nylon meshes. Proc. Natl. Acad. Sci. USA 76: 283–287.
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Suzuki, M., Ichikawa, K., Sakoda, A. et al. Long-term culture of primary rat hepatocytes with high albumin secretion using membrane-supported collagen sandwich. Cytotechnology 11, 213–218 (1993). https://doi.org/10.1007/BF00749872
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DOI: https://doi.org/10.1007/BF00749872