Neural crest neuroblasts can colonise aganglionic and ganglionic gut in vivo

Introduction: Neural crest (NC) cells differentiate in vitro into neuroblasts, precursors of the enteric nervous system (ENS), when stimulated by specific agents. We developed a study aimed at establishing whether NC-derived neuroblasts can survive and colonise in vivo when injected into a recipient mouse gut. Materials and Methods: The neuroblast precursors of the ENS were obtained from the vagal portion of the neural tubes of 296 CD-1 and Gtrosa26 mouse embryos. The embryonic cells of Gtrosa26 mice are identifiable through beta-galactosidase activity which allows recognition by blue staining. The host used in this study was the Dom/⁺ mouse, an animal model for Hirschsprung's disease (aganglionic megacolon). Dom/⁺ mouse pups (n = 43) received NC-derived cells inoculated into the seromuscular layer of the gut (33/43) or directly into the peritoneal abdominal cavity (10/ 43). Results: All Dom/⁺ mice survived the procedure and were sacrificed after 7 or 14 days. Histochemical staining detected implanted cells in all mice. These showed specific myenteric colonisation into the aganglionic and ganglionic gut. Conclusion: The striking result of this study was the specific tropism of the injected NC-derived cells to target sites under the action of unknown chemotactic agents. This experimental procedure might represent a possible treatment option for specific forms of human ENS anomaly such as total intestinal aganglionosis.