Evom endohm 12chamber

The Caco-2 (ATCC^® HTB-37™) human colorectal cancer cell line is a continuous line of heterogeneous human epithelial colorectal adenocarcinoma cells. Caco-2 cells differentiate into enterocytes on collagen-coated Transwell^® polyester membrane inserts (Corning 3640). Enterocytes are characterised by tight junctions, microvilli, and a number of specific enzymes and transporters [49 (link)]. Caco-2 cells were grown in EMEM medium supplemented with 10% heat-inactivated foetal bovine serum (FBS), 2 mM L-glutamine, 0.1 mM non-essential amino acids, 100 U/mL penicillin, and 100 µg/mL streptomycin, and maintained at 37 °C in a humidified environment with 5% CO₂. Before the experiments, cells were seeded at 90,000 cells/cm² onto collagen-coated Transwell^® membrane inserts and maintained in culture for 21 days in order to reach a reliable intestinal barrier. The transepithelial electrical resistance (TEER) was measured every 7 days by means of an EVOM EndOhm-12chamber (World Precision Instruments, Sarasota, FL, USA), as a parameter of barrier attainment. The medium was replaced every 3 days in the basolateral and the apical chambers, which represent the circulatory and luminal poles of the intestinal epithelium, respectively.

The anti-inflammatory capacity experiments were performed in an in vitro intestinal barrier model since it represents the most suitable condition to study the gut maintenance and defense to inflammatory mediators and the modulation of intestinal epithelial permeability. Caco-2 cellswere seeded onto collagen coated Transwell^® polyester membrane inserts (Corning 3640) and maintained at 37 °C in a 5% CO₂ atmosphere for 21 days, in order to reach the proper differentiation in a polarized epithelial cell monolayer. The basolateral and the apical sides of the insert represent the circulatory and luminal poles of the intestinal epithelium, respectively. Medium was replaced both in apical and basolateral chambers every 3 days, and Transendothelial Electrical Resistance (TEER) was measured every 7 days by means of EVOM EndOhm-12 chamber (World Precision Instruments, Sarasota, FL, USA), to check the formation of a reliable intestinal barrier.
TEER was evaluated as a parameter of barrier functionality after inflammation in cells pretreated or not with cinnamon digested extract. TEER measures were performed before and after the 24 h-inflammatory treatment for every insert, and the ΔTEER [Δ (Ω·cm²)] between the two measures was calculated and analyzed within the groups.