The development of a method for the preparation of rat intestinal epithelial cell primary cultures.
Affiliation
Department of Epithelial Biology, Paterson Institute for Cancer Research, Christie Hospital and Holt Radium Institute, Withington, Manchester, UK.Issue Date
1992-01
Metadata
Show full item recordAbstract
We describe a reproducible method for growing small intestinal epithelium (derived from the suckling rat intestine) in short-term (primary) cultures. Optimal culture conditions were determined by quantitative assays of proliferation (i.e. changes in cellularity and DNA synthesis). Isolation of the epithelia and, significantly, preservation of its three-dimensional integrity was achieved using a collagenase/dispase digestion technique. Purification of the epithelium was also facilitated by the use of a simple differential sedimentation method. The results presented below support the idea that proliferation of normal gut epithelium ex vivo is initially dependent upon the maintenance of the structural integrity of this tissue and upon factors produced by heterologous mesenchymal cells. Proliferation in vitro was also critically dependent upon the quality of the medium and constituents used. Cultures reached confluence within 10-14 days and consisted of epithelial colonies together with varying amounts of smooth-muscle-like cells. Cultures have been maintained for periods up to one month, but the longer-term potential for growth by sub-culturing has not been examined. Strategies for reducing the proliferation of these non-epithelial cells are also described.Citation
The development of a method for the preparation of rat intestinal epithelial cell primary cultures. 1992, 101 ( Pt 1):219-31 J. Cell. Sci.Journal
Journal of Cell SciencePubMed ID
1569126Type
ArticleLanguage
enISSN
0021-9533Related articles
- [Cultures of the mucosa of the small intestine].
- Authors: Rampal P, Delmont J
- Issue date: 1975 Mar
- Establishment of conditionally immortalized epithelial cell lines from both colon and small intestine of adult H-2Kb-tsA58 transgenic mice.
- Authors: Whitehead RH, VanEeden PE, Noble MD, Ataliotis P, Jat PS
- Issue date: 1993 Jan 15
- [Enzymatic-transport system of epithelial cells, stroma, and the muscle-serous layer of the rat small intestine].
- Authors: Gromova LV, Gusev SA, Egorova VV, Iezuitova NN, Nikitina AA, Timofeeva NM, Tsvetkova VA, Ugolev AM
- Issue date: 1991
- Effects of extracellular iron concentration on calcium absorption and relationship between Ca2+ and cell apoptosis in Caco-2 cells.
- Authors: Wang L, Li Q, Duan XL, Chang YZ
- Issue date: 2005 May 21
- Ex vivo gut culture for studying differentiation and migration of small intestinal epithelial cells.
- Authors: Sun X, Fu X, Du M, Zhu MJ
- Issue date: 2018 Apr