title: Good Caco-2 cell culture practices.
abstract: The human Caco-2 cells differentiate spontaneously in culture forming monolayers of mature intestinal enterocytes which have been used as a model of the intestinal barrier for in vitro toxicology studies. Reproducibility problems often reported in literature have been generally ascribed to different culture-related conditions, such as the type of animal serum used, the supplements added to the culture media, the passage number and the source of cell clones. The Caco-2 cell culture protocol here described has been recently optimized in our laboratory, producing a homogeneous and highly polarized monolayer of cells which display many of the characteristics of the intestinal enterocytes. This protocol differs from standard protocols mainly because Caco-2 cells are subcultured when they reach just 50% of confluence, instead of 80%, retaining a high proliferation potential. When this cell population is seeded at high density on filter inserts differentiates almost synchronously and much more homogenously.
authors.affiliation: CNR, Institute of Cell Biology and Neurobiology, Rome, Italy.
authors.forename: Manuela
authors.initials: M
authors.lastname: Natoli
authors.forename: Bruno D
authors.initials: BD
authors.lastname: Leoni
authors.forename: Igea
authors.initials: I
authors.lastname: D'Agnano
authors.forename: Flavia
authors.initials: F
authors.lastname: Zucco
authors.forename: Armando
authors.initials: A
authors.lastname: Felsani
mesh_terms: D000818:Animals; D018938:Caco-2 Cells; D018929:Cell Culture Techniques; D002454:Cell Differentiation; D049109:Cell Proliferation; D002478:Cells, Cultured; D003470:Culture Media; D020895:Enterocytes / Q000378:metabolism*; D006801:Humans; D008954:Models, Biological; D015203:Reproducibility of Results; D014116:Toxicology / Q000379:methods
chemical_list: D003470:Culture Media