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, remain unclear. Therefore, we aimed to examine ENSC integration using an adapted ex vivo organotypic culture system. Donor ENSC were obtained from Wnt1cre/+;R26RYFP/YFP mice allowing specific labelling, selection and fate-mapping of cells. YFP+ neurospheres were transplanted to C57BL6/J (6–8-week-old) colonic tissue and maintained in organotypic culture for up to 21 days. We analysed and quantified donor cell integration within recipient tissues at 7, 14 and 21 days, along with assessing the structural and molecular consequences of ENSC integration. We found that organotypically cultured tissues were well preserved up to 21-days in ex vivo culture, which allowed for assessment of donor cell integration after transplantation. Donor ENSC-derived cells integrated across the colonic wall in a dynamic fashion, across a three-week period. Following transplantation, donor cells displayed two integrative patterns; longitudinal migration and medial invasion which allowed donor cells to populate colonic tissue. Moreover, significant remodelling of the intestinal ECM and musculature occurred upon transplantation, to facilitate donor cell integration within endogenous enteric ganglia. These results provide critical evidence on the timescale and mechanisms, which regulate donor ENSC integration, within recipient gut tissue, which are important considerations in the future clinical translation of stem cell therapies for enteric disease.
Highlights
Enteric neural stem cells (ENSC) have been identified as a possible treatment for enteric neuropathies
We further demonstrate that such integration requires the dynamic remodelling of collagen components within the extracellular matrix (ECM), and tissue architecture, as donor cells migrate across the gut wall
To assess the integration of ENSC, within intestinal segments, we developed an ex vivo organotypic culture model, based upon methodology previously used for enteric nervous system (ENS) imaging[20,21], in which colonic tissues could be maintained in situ with limited contractile forces (Fig. 1A–D), which typically lead to tissue damage when pinned
Summary
Enteric neural stem cells (ENSC) have been identified as a possible treatment for enteric neuropathies. 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. Significant remodelling of the intestinal ECM and musculature occurred upon transplantation, to facilitate donor cell integration within endogenous enteric ganglia These results provide critical evidence on the timescale and mechanisms, which regulate donor ENSC integration, within recipient gut tissue, which are important considerations in the future clinical translation of stem cell therapies for enteric disease. Proof-of-principle studies have established the potential for in vivo transplantation of ENSCderived neurons in wild-type[14,15,16] and dysmotile transgenic tissues[17,18] These studies have shown the successful long-term engraftment of ENSC and their derivatives within the colonic muscularis. Donor-derived neurons have been observed at considerable distances from the presumptive site of transplantation, and appear able to migrate through the muscularis to reside within endogenous ganglia structures at the level of the myenteric plexus
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Free) 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a call
