Term Epithelial Cultures

Epithelial lines can be obtained by maintenance of the primary cultures of adult hepatocytes beyond the initial period of deterioration. Without special precautions, few of the cultures will be free of fibroblasts. A variety of procedures can minimize this problem, including treatment with 1 fxM dexa-methasone. Exposure to dexamethasone for 4 days followed by a switch back to medium E containing 10% FBS will stimulate growth of epithelial colonies. These may be identified and marked and the remainder of the culture 

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The ability of both hepatocyte primary cultures and long-term epithelial cultures to metabolize a spectrum of activation-dependent carcinogens is demonstrated indirectly by the production of transformation, DNA damage,DNA repair (Section 6), and mutagenesis (Sections 7 and 8) in these cultures. 

Williams, G. M., and Gunn, J. M., 1974, Long-term cell culture of adult rat liver epithelial cells, Exp. Cell Res. 89 139-142. 

Long-term epithelial liver cultures maintain a significant level of aryl hydrocarbon hydroxylase. However, substantial heterogeneity in both... 

Some laboratories have found an alternative to the short-term cultures by using cell lines other than Caco-2 cells. The most popular of these is Madin-Darby canine kidney (MDCK) cells, an epithelial cell line from the dog kidney. MDCK cells have been suggested to perform as well as Caco-2 cells in studies of passive drug permeability [56]. These cells have also been used to optimise the conditions for studies of low-solubility drugs [53]. However, as noted previously, the active transport processes of this cell line can be quite different to those of Caco-2 cells [28-30], Another cell line that only requires short-term culture is 2/4/A1, which is a conditionally immortalised rat intestinal epithelial cell line [86]. The 2/4/A1 cell line is discussed in Section 4.3.2.2 below. 

Guillouzo et al. (1988) developed a coculture system of rat or human hepatocytes with rat liver epithelial cells that maintains the hepatocytes in a differentiated state for extended periods of time, thereby allowing studies involving chronic treatment with the test substance to be conducted. Primary cultures of hepatocytes can therefore provide a useful model for short- and long-term studies involving the safety assessment of xenobiotics. 

V. E. Steele and J. T. Arnold. Isolation and long-term culture of rat, rabbit and human nasal turbinate epithelial cells. In Vitro Cell Dev Biol 21 671-687 (1985). 

D. C. Gruenert, C. B. Basbaum, and J. H. Widdicombe. Long-term culture of normal and cystic fibrosis epithelial cells grown under serum-free condition. In Vitro Cell Dev Biol 26 411-418 (1990). 

Barrett, J.C., Gray, T.E., Mass, M.J., and Thomassen, D.G. (1982). A quantitative, clonal assay for carcinogen-induced alterations of respiratory epithelial cells in culture, in Application of Short-term Bioassays in the Analysis of Complex Environmental Mixtures, Waters, M. and Sandu, S., Eds. (Plenum Press, New Y>rk). 

Wiszniewski L, Jornot L, Dudez T et al (2006) Long-term cultures of polarized airway epithelial cells from patients with cystic fibrosis. Am J Respir Cell Mol Biol 34(1) 39 8... 

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