Recreation in vitro of human intestinal tissue using advanced cell culture technology

Abstract

The small intestine has the main function of absorbing nutrients and plays a prominent role in the normal function of the body. In vitro models have been developed to study the process of molecular transport across the intestinal epithelium as an entry point into the body. This is of particular importance in industry and the ability to perform permeability assays. Conventional intestinal in vitro model consists of a single monolayer of epithelial cells, such as the Caco-2 lineage, cultured on a permeable and flat membrane. However, this system has many limitations. It has been demonstrated that the barrier function in the Caco-2 monolayer does not really represent the features of the native small intestinal tissue. Intestinal tissue consists of an integrated system where multiple different cell types interact each other to maintain and perform their function. In this project the effect of coculturing Caco-2 intestinal epithelial cells with fibroblasts was investigated to more closely mimic the structure and function of native tissue and to create a model of the intestinal mucosa. Structural and functional changes were examined during the maturation of Caco-2 cells when they were grown with paracrine factors derived from fibroblasts or in direct co-culture with fibroblasts in a novel 3D cell culture scaffold.
These data indicate that human colon fibroblasts (CCD-18co) and neonatal human dermal fibroblasts (HDFn) produce signals that influenced the Caco-2 barrier structure. It was identified that this change was most likely due to a reduction in junctional complexes between the epithelial cells which are important proteins in the control of paracellular transport. It was demonstrated that this novel 3D intestinal model offers a promising system with greater anatomical and physiological relevance.