Epithelial Cell Culture Protocols by Clare Wise

By Clare Wise

Well-versed experimenters and medical researchers percentage their top equipment for setting up and preserving epithelial mobilephone cultures, for interpreting and learning their features, and for utilizing them to establish versions of severe organic structures. The emphasis is at the research and evaluation of epithelial cells, for instance by means of taking a look at apoptosis and integrins, or by means of measuring membrane capacitance and confluence. additionally defined in step by step element are co-culture ideas beneficial in constructing types for investigating many various in vitro platforms, together with the blood-brain barrier, drug uptake, and the interplay of epithelial cells with micro organism. Epithelial mobile tradition Protocols deals a step by step consultant towards a deeper figuring out of mobile and molecular mechanisms, in addition to a suite of strong suggestions for in particular comparing the character and behaviour of epithelial cells.

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188: Epithelial Cell Culture Protocols Edited by: C. , Totowa, NJ 37 38 Sirica hyperplastic biliary epithelium (27,28), and the third developed to establish a novel rat cholangiocarcinoma cell culture model (29). The first protocol that will be described represents a more recent modification developed by us (27) of our initial 1985 method (1) used to isolate and culture hyperplastic biliary epithelial cells from 6- to 15-wk bile duct-ligated rat liver. 2. details our method for isolating and culturing nontransformed hyperplastic biliary epithelial cells from the livers of 7-wk bile duct-ligated/6-wk furan-treated rats (28).

Pieces which do not adhere to the bottom after 3 d should be removed. 19. Monitor the progress of the cultures by observing them using an inverted microscope. After 2–3 d, epithelial cells should be seen expanding from the thymic pieces, forming islets with a few single cells between the islets. The islets expand and may grow to confluence within 2–3 wk. Thereafter, expansion usually stops, but cultures are viable for a few weeks more. 20. In the second or third week of culture, cells may be transferred to new Vitrogencoated culture chambers by trypsinization.

14. , Benedict, C. , Picciano, P. , and Cohen, S. (1987) Substrate requirements for the isolation and purification of thymic epithelial cells. J. Exp. Pathol. 3, 251–258. 15. Nieburgs, A. , Picciano, P. , Korn, J. , and Cohen, S. (1985) In vitro growth and maintainance of two morphologically distinct populations of thymic epithelial cells. Cell. Immunol. 90, 439–450. 16. , Petersen, O. , and van Deurs, B. (1990) Short-term cultivation of murine thymic epithelial cells in a growth factor defined serum-free medium.

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