HUMAN INTESTINAL SPHEROIDS AS A MODEL SYSTEM FOR STUDYING ABC TRANSPORTERS

GOMEZ BURICH A.G; ZECCHINATI F; RICARDI L.L; ARANA M.R; CEBALLOS, MARÍA PAULA; PALMA NICOLÁS FRANCISCO; PERDOMO V.G; VILLANUEVA S.S.M

Buenos Aires

Congreso; Reunión Anual de Sociedades de Biociencias (SAIC SAFIS ALACF); 2024

Sociedad Argentina de Investigación Clínica (SAIC)

The human Caco-2 monolayer culture is a valuable in vitro model for studying intestinal barrier function, with a particular focus on the role of ATP-binding cassette (ABC) proteins in mediating drug transport. However, 3D in vitro models, which better replicate the complex in vivo environment due to their ability to capture tissue-specific architecture and cell-cell interactions, have become increasingly important. Even so, these models still require characterization. Objectives: To generate a Caco-2 spheroid model using the Liquid Overlay technique and evaluate the ABCC2 and ABCB1 protein expression. Methods: Caco-2 spheroids were formed by seeding 10.000 cells/well onto 96-well plates coated with 1.5% agarose. After 4 and 7 days of culture, spheroid diameter was measured, and cell viability was assessed by MTT assay and LDH activity. ABCC2 and ABCB1 protein expression was analyzed using Western blotting (WB). Statistical analyses were performed using the t-test. Results: Caco-2 spheroids exhibited reproducible diameter of 866±34 μm after 4 days of culture and 1041±65 μm after 7 days, with a statistically significant difference between the two-time points (p