EVALUATION OF SURFACES FOR INTESTINAL EPITHELIAL CELL CULTURE

BRUN ANTONIO; ENRIQUE CAVIEDES-VIDAL

San Juan

Congreso; SECOND JOINT MEETING OF THE BIOLOGY SOCIETIES FROM ARGENTINA; 2011

Sociedad Argentina de Biología, Sociedad de Biología de Cuyo, Sociedad de Biología de Córdoba, Universidad Católica de Cuyo, Universidad Nacional de San Juan

The intestinal epithelium is rapidly renewed through a process of differentiation of stem cells that migrate from the crypt along the major axis of the villi. During this process there have been identified several factors influencing this phenomenon, including the extracellular matrix. The study aims to assess different surfaces for intestinal epithelial cell culture. Studied surfaces were polyethylene terephthalate (PET) from Nunc and Grenier Bio-One, collagen type I (Invitrogen) and Matrix cell (BDBioscience). The primary culture was performed according to the protocol develop by Campbell et al. 2008. The culture medium used, D-MEM supplemented with 2.5% FBS, insulin 10 mg/ml, transferrin 5.5 mg/ml, selenium 0.67 mg/ml, porcine mucosal heparine 50 μg/ml, penicillin 50 U/ml and streptomycin 50μg/ml, was incubated at 37ºC and 5% CO2 . The medium was renewed every other day and growth evaluation was performed daily using an inverted microscope. Dissimilar results were obtained according to the surface to be evaluated. Cell growth was apparent on PET Nunc surface. We obtained an homogeneous monolayer of cells with epithelial morphology that lasted up to 22 days. On the surface PET Grenier Bio-One, cell proliferation was practically nil. Regarding biological surfaces, in both were cell growth, but only achive to obtain a homogeneous monolayer with epithelial morphology alone on collagen. We obtained better adherence of cells in the first 24 hours of culture in biological surfaces and longer confluence time in the plastic surface. Subcultures were not positive in none of the tested surfaces. Conclusion: Collagen surface was found to be most suitable for the proliferation of intestinal epithelial cells. Funded by PICT97-01320 to EC-V