New methodological approach to induce a differentiation phenotype in Caco-2 cells prior to post-confluence stage
Ferraretto, A.; Gravaghi, C.; Donetti, E.; Cosentino, S.; Donida, B.M.; Bedoni, M.; Lombardi, G.; Fiorilli, A.; Tettamanti, G.
Anticancer Research 27(6b): 3919-3925
2007
ISSN/ISBN: 0250-7005 PMID: 18225551 Document Number: 605940
Various differentiation-inducing agents or harvesting of spontaneously late post-confluence cultures have been used to differentiate the human colon carcinoma Caco-2 cell line. We report a new procedure to generate pre-confluent subcultures of Caco-2 population at various stages of differentiation without altering culture conditions. Ultrastructural analysis, cell proliferation activity and biochemical markers of differentiation were evaluated at different passages. Subcultures of Caco-2 cells at pre-confluence, exhibiting progressive acquisition of a more benign differentiation phenotype, were generated. Early passages of Caco-2 cells showed a well-developed brush border and incomplete junctional apparatus; subsequent subcultures yielded cell populations with well-developed junctions similar to those of small intestinal cells. These culture conditions represent a new versatile model not only to progressively induce the differentiation program in Caco-2 cells at pre-confluence without changes of culture media, but also to explore mechanistic modes of drug transport and tumor development.