Isolation, Expansion and Transplantation of Postnatal Murine Progenitor Cells of the Enteric Nervous System

Abstract

Neural stem or progenitor cells have been proposed to restore gastrointestinal function in patients suffering from congenital or acquired defects of the enteric nervous system. Various, mainly embryonic cell sources have been identified for this purpose. However, immunological and ethical issues make a postnatal cell based therapy desirable. We therefore evaluated and quantified the potential of progenitor cells of the postnatal murine enteric nervous system to give rise to neurons and glial cells in vitro. Electrophysiological analysis and BrdU uptake studies provided direct evidence that generated neurons derive from expanded cells in vitro. Transplantation of isolated and expanded postnatal progenitor cells into the distal colon of adult mice demonstrated cell survival for 12 weeks (end of study). Implanted cells migrated within the gut wall and differentiated into neurons and glial cells, both of which were shown to derive from proliferated cells by BrdU uptake. This study indicates that progenitor cells isolated from the postnatal enteric nervous system might have the potential to serve as a source for a cell based therapy for neurogastrointestinal motility disorders. However, further studies are necessary to provide evidence that the generated cells are capable to positively influence the motility of the diseased gastrointestinal tract.

Delivery of enteric neural progenitors with 5-HT4 agonist-loaded nanoparticles and thermosensitive hydrogel enhances cell proliferation and differentiation following transplantation in vivo

“…The success of this approach can be attributed to sustained release of 5-HT 4 agonist from liposomal nanoparticles, maintenance of spatial proximity of cells and particles when contained within the hydrogel, and the immune isolation afforded by hydrogel encapsulation. These findings support this novel approach that enhances the otherwise limited ENSC proliferation and differentiation previously reported [9, 10] and has the potential to significantly enhance the success of cell transplantation for treating neurointestinal diseases.…”

“…Encapsulation of the 5-HT 4 receptor agonist, RS67506, within liposomal nanoparticles effectively maintains this effect following ENSC transplantation in vivo . This represents a novel approach that enhances the otherwise limited proliferation and differentiation potential of ENSCs observed in prior transplantation studies [6, 9, 10]. …”

“…However, it has also been shown that functional improvement lasts only a short period (up to 1 week) [10, 11] and the migration and differentiation potency of the transplanted cells are limited [9, 12]. Previous studies have manipulated the stem cells using lentiviral vectors [51] or microenvironment modification by local injection of an anti-inflammatory drug [52] in order to enhance cell survival following transplantation.…”

“…A number of recent studies have shown that ENS stem/progenitor cells (ENSCs) can be isolated, expanded in culture, and transplanted into intestinal tissue in vitro [4-6] and in vivo [7-9], including into animal models of enteric neuropathies [10-12]. However, the capacity of these cells to proliferate and migrate has been limited [12, 13], and may therefore be insufficient to restore gut function, emphasizing the need to engineer the cells and/or their microenvironment, and to optimize cell delivery methods, in order to maximize the success of cell replacement therapy.…”

Enteric neural progenitors are more efficient than brain-derived progenitors at generating neurons in the colon

“…A variety of sources of stem/progenitor cells has been examined, including the ENS, the central nervous system (CNS), and skin, as well as embryonic and induced pluripotent stem cells (16,21,27,28,35,37,61,67). It is essential that the source of stem/progenitor cells can be sufficiently expanded in vitro and subsequently generate mature, functional neurons of the appropriate phenotype.…”