By Clare Wise
There were major advances in examine concerning the isolation and tradition of epithelial cells some time past decade, and lots of new thoughts were built. Monolayer cultures can be utilized to judge the character and behaviour of cells, whereas using epithelial cells in version platforms has allowed a deeper realizing of mobile and molecular mechanisms and interactions. the purpose of this e-book is to supply a accomplished, step by step consultant to many innovations for epithelial mobilephone tradition, combining in a single quantity the regularly used protocols in addition to many who are extra speci- ized. Epithelial telephone tradition Protocols may help people who are new to this box and wish to profit the fundamental tradition suggestions, in addition to these desiring to take advantage of extra large ranging and particular protocols. it may be an invaluable source by itself, and likewise supplement the opposite volumes which have been written approximately cellphone tradition within the tools in Molecular Biology sequence. Epithelial cellphone tradition Protocols covers a large choice of protocols, regularly aimed toward the researcher, but additionally a couple of aimed toward clinicians. The est- lishment and upkeep of fundamental cultures derived from many alternative tissues and assorted species is roofed. specific emphasis has been put on protocols had to additional learn and determine epithelial cells, for instance, by means of taking a look at apoptosis and integrins and through measuring membrane capa- tance and confluence. utilizing varied co-culture innovations, it's attainable additionally to boost versions to enquire many various structures in vitro.
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120, 67–78. 2. , Faris, R. , and Hixson, D. C. (1993) Long-term culture and characteristics of normal rat liver bile duct epithelial cells. Gastroenterology 104, 840–852. 3. , and Toyota, T. (1995) Confluent monolayers of bile duct epithelial cells with tight junctions. Hepatology 22, 153–159. 4. Vroman, B. and LaRusso, N. F. (1996) Development and characterization of polarized primary cultures of rat intrahepatic bile duct epithelial cells. Lab. Invest. 74, 303–313. 5. Sirica, A. E. (1992) Biology of biliary epithelial cells, p.
5. , Jenkinson, E. , Moore, N. , and Owen, J. J. T. (1993) MHC class II-positive epithelium and mesenchyme cells are both required for T-cell development in the thymus. Nature 362, 70–73. 6. Ernst, B. , Surh, C. , and Sprent, J. (1996) Bone marrow-derived cells fail to induce positive selection in thymus reaggregation cultures. J. Exp. Med. 183, 1235–1240. 7. Ropke, C. (1997) Thymic epithelial cell culture. Microsc. Res. Tech. 38, 276–286. 8. , and Sato, G. (1980) Methods for growth of cultured cells in serumfree medium.
Collagenase (type I) (Sigma). 95% Oxygen/5% carbon dioxide. DNase I (Sigma). Bile Duct Cell Culture 39 Fig. 1. Examples of fine-tooth combs used to effectively separate intact hyperplastic biliary tissue from hepatic parenchyma following perfusion of bile duct-ligated rat liver in situ with collagenase-type 1. 10. 1 µM insulin, 2 mM L-glutamine, 3% fetal bovine serum (FBS) (Sigma), 100,000 U/L penicillin G, and 100 mg/L streptomycin sulfate, 360,000 U/L collagenase (type I), 130,000 U/L DNase I, 700,000 U/L hyaluronidase (type III) (Sigma), and 100 mg/L soybean trypsin inhibitor (type I-S) (Sigma).
