Abstract
Reliable toxicological risk assessment (RA) is crucial for the safety of the public. Caco2 - an adenocarcinoma cell line - is commonly used when intestinal toxicity screening.
Nevertheless, Caco2 monoculture does not reflect upon the complexity of the intestinal tissue. To address this shortcoming, we have employed an intestinal cell line HT29 together with Caco2 in a co-culture model.
Initially, a suitable ratio of the two cell lines was investigated in terms of alkaline phosphatase activity (ALP) (days 3, 14 and 21 post-seeding) and mucin production (day 21 post-seeding) on tissue culture plastic (TCP). Next, selected co-culture (Caco2:HT29 7:3) was cultured on inserts with a nanofibrous scaffold (a mixture of poly-ε-caprolactone and cellulose acetate) in order to mimic a 3D in vivo architecture.
In this case, the preservation of both cell lines was verified by staining HT29 with a Cell Tracker and subsequent confocal microscopy visualization. Finally, in order to determine a suitable post-seeding time frame for the toxicological screening, the β-actin was visualized using confocal microscopy (days 1, 4 and 7 post-seeding) and ALP activity was measured (days 3, 7, 14 and 21 post-seeding).