By Clare Wise
Well-versed experimenters and scientific researchers proportion their most sensible tools for constructing and holding epithelial mobile cultures, for interpreting and learning their features, and for utilizing them to establish types of serious organic platforms. The emphasis is at the research and evaluation of epithelial cells, for instance through apoptosis and integrins, or by means of measuring membrane capacitance and confluence. additionally defined in step by step aspect are co-culture ideas worthwhile in constructing types for investigating many alternative in vitro structures, together with the blood-brain barrier, drug uptake, and the interplay of epithelial cells with micro organism. Epithelial mobilephone tradition Protocols deals a step by step consultant towards a deeper knowing of mobile and molecular mechanisms, in addition to a suite of sturdy thoughts for in particular comparing the character and behaviour of epithelial cells.
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Additional info for Epithelial Cell Culture Protocols
Like rat hyperplastic biliary epithelium in gel culture, the cultured rat cholangiocarcinoma cells exhibit strong cytoplasmic immunoreactivity for cytokeratin 19 (Figs. 3F, G), but unlike hyperplastic biliary epithelium, the cultured cholangiocarcinoma cells, similar to neoplastic glandular epithelium of the parent tumor, give a positive histochemical reaction for mucin (29) (Fig. 3H). Utilizing differential cell harvesting, involving selective trypsinzation and serial passaging of cultured rat cholangiocarcinoma cells, we established a novel rat cholangiocarcinoma epithelial cell line designated C611B (29).
8. Human thymic fragments may be stored overnight in medium at 37°C or at 4°C before being put to culture, or they may be frozen in liquid nitrogen in medium containing 10% dimethyl sulfoxide (DMSO) and 10% serum. However, seeding efficiency will be inferior to the one obtained using fresh tissue. 9. After trypsinization, cultured human epithelial cells may be kept in liquid nitrogen in 10% DMSO plus 10% serum for a prolonged time and are easily brought to culture by seeding about 105 cells/mL.
3. In the sterile hood, transfer the tissue into a glass dish and wash several times by flooding with PBS. 4. Select suitable pieces free of connective tissue and blood and cut them off using surgical knives. 5. Transfer the pieces to another dish, cut them into 1–2-mm fragments, and transfer these into a 12-mL tube using a finnpipet. The plastic tip of the latter is cut obliquely to allow entrance of the thymic pieces. 6. Wash the pieces twice with 10 mL medium. 7. Aspirate the medium from the tube and add 10 mL of sterile filtered collagenase/ dispase/DNase solution.