Three-dimensional co-culture model of enterocytes and primary enteric neuronal tissue

The aim of this study was to establish a three-dimensional model of the innervated mucosal barrier using a co-culture of an enterocyte cell line and enteric glial and nerve cells. Such a model might form the basis for further studies of interactions between the single compartments of the bowel wall, as well as of extrinsic influences on intestinal development and plasticity.

Isolated and dissociated myenteric plexus was resuspended in either collagen or extracellular matrix (ECM) solutions. After incubation at 37 degrees C the solution gelled and formed stable plugs where neurons and glial cells reaggregated to form secondary neuronal networks. HT-29-enterocytes were seeded on top of the gels either immediately (collagen, ECM), or after adding a thin layer of collagen II (ECM).

While the neuronal tissue formed complex networks within the gel, the enterocytes on top of the gels grew differently depending on the substrate and innervation. So enterocytes on ECM gels did not grow to confluence, while on collagen gels or on ECM plus collagen larger patches and increasing confluence could be observed. In general HT-29 grew better on innervated gels than on gels with no neuronal tissue.

With the presented model of different compartments of the bowel wall, various parameters of intercellular dependencies and influences can be observed in vitro. Moreover, the first results are also steps towards developing an innervated gut wall in vitro which might be able to restore functional capacity in infants with short bowel syndrome or other disorders that severely impair bowel function.