Transplantation of amniotic fluid-derived neural stem cells as a potential novel therapy for Hirschsprung's disease

Abstract

Background/purpose: We have previously shown that embryonic enteric neural stem cells (NSCs) isolated from the intestine colonize aganglionic intestine upon transplantation, but posttransplantation cell survival limits efficacy. The aims of this study were to investigate whether transplantation of amniotic fluid (AF)-derived NSCs could improve survival of the engrafted cells and promote functional recovery of the diseased colon.

Methods: AF cells were induced into NSCs with neurogenic medium, and further differentiated into neurons and glial cells. Ednrb knockout mice received an intestinal intramuscular injection of 20,000 AF-derived NSCs into the aganglionic colon. Engrafted cells were visualized and characterized by immunohistochemistry for GFP, neuronal, and glial cell markers. Colonic motility was quantified by colonic bead expulsion time.

Results: AF-derived NSCs had increased expression levels of the NSC marker Nestin and the glial cell marker GFAP compared to enteric NSCs. Transplanted AF-derived NSCs had decreased apoptosis and increased survival compared to enteric NSCs. Colonic motility was significantly improved in Ednrb knockout mice transplanted with AF-derived NSCs, as demonstrated by significantly decreased colonic bead expulsion time.

Conclusion: AF-derived NSCs have enhanced survival upon transplantation into a defective enteric nervous system. Transplantation of AF-derived NSCs may represent a potential novel future therapy for the treatment of Hirschsprung's disease.

Keywords: Amniotic fluid; Apoptosis; Hirschsprung's disease; Neural stem cell; Transplantation.