Dynamic integration of enteric neural stem cells in ex vivo organotypic colon cultures

Abstract

Enteric neural stem cells (ENSC) have been identified as a possible treatment for enteric neuropathies. After in vivo transplantation, ENSC and their derivatives have been shown to engraft within colonic tissue, migrate and populate endogenous ganglia, and functionally integrate with the enteric nervous system. However, the mechanisms underlying the integration of donor ENSC, in recipient tissues, remains unclear. Here, using a modified ex vivo organotypic culture system we show that donor ENSC-derived cells integrate across the colonic wall in a dynamic fashion, across a three-week period. We further show that donor cells display two integrative patterns; longitudinal migration and medial invasion which allow donor cells to populate colonic tissue. Moreover, we demonstrate that significant remodelling of the intestinal ECM, and musculature, occurs upon transplantation to facilitate donor cell integration. Thus, our results provide critical evidence on the timescale, and mechanisms, which regulate donor ENSC integration within recipient gut tissue.