Details

Autor(en) / Beteiligte

• Kaeffer, B
• Briollais, S

Titel

Primary Culture of Colonocytes in Rotating Bioreactor

Ist Teil von

• In vitro cellular & developmental biology. Animal, 1998-09, Vol.34 (8), p.622-625

Ort / Verlag

Germany: Society for In Vitro Biology

Erscheinungsjahr

1998

Quelle

SpringerLINK Contemporary (Konsortium Baden-Württemberg)

Beschreibungen/Notizen

• Development of simple methodologies to obtain cultures of normal epithelial cells is especially relevant for study of the influences of dietary and gut growth factors and prevention of cell death during organ transplantation or inflammatory processes. Rotating bioreactors allowing three-dimensional cocultures of normal or cancerous cells under conditions of simulated microgravity are now used in a wide range of tissue culture applications, including studies on tissue morphogenesis. The three-dimensional cultures of human small intestinal cells in rotating-wall vessels (RWV) lead to the obtainment of small clumps of fully differentiated tissues. Recently, we have described a short-term primary culture method optimizing colonic cell adhesion to plastic substratum or to cytodex-3 microcarriers after exposure to sodium orthovanadate in order to assay the in vitro effects of food oligosaccharides on colon morphogenesis. The rationale behind these experiments has been that vanadate (VO sub(4) super(3-)) is a general inhibitor of protein-tyrosine phosphatases (PTP) acting as a phosphate analog by adopting a trigonal bipyramidal structure; the mechanism of inhibition and its full reversibility in an enzyme assay has been recently described. Under physiological conditions sodium orthovanadate mimics insulin and has the capacity to inhibit PTP, slowing programmed cell death processes as well as to reducing the density-dependent difference in tyrosine phosphorylation governing cell survival at high density. The consequence of an exposure to sodium orthovanadate for the survival and growth of tissue-like masses with typical colonocytes could be directly studied in a rotary cell culture system, paving the way for development of more complex experiments on the regulation of colonocyte growth and differentiation by transforming growth factor alpha (TGF-a). In this study we report experiments performed in RWV to maintain rat primary cultures for 22 d after removing sodium orthovanadate from tissue culture medium. The toxicity of sodium orthovanadate and its capacity to delay programmed cell death were quantitatively studied on IEC-6 and Caco-2 cell lines.