Differentiation of Gastrointestinal Cell Lines by Culture in Semi-wet Interface

Abstract

Epithelial cells grown in vitro provide opportunities to elucidate cellular mechanisms in response to chemical, viral, or bacterial agents in isolation from the effects of other bodily functions, such as hormonal and immune responses. However, cells that do not form a tight epithelium, polarize or secrete mucins lack some of the important protection mechanisms intrinsic to epithelial cells in vivo, increasing their susceptibility to external agents, and exposing basolateral targets for interactions that may not occur in vivo. Here, we present a method that transforms some epithelial cell lines into mucin secreting polarized epithelial surfaces with high transepithelial resistance: the cells are cultured on semi-permeable membranes in differentiation medium for the first 6 days, followed by culture under semi-wet interface with mechanical stimulation for 22 days. The procedure can be performed using standard laboratory reagents and equipment. A description on how to fix and paraffin embed these in vitro mucosal membranes for histology purposes is also included.