Stem cell therapy offers the potential of rebuilding the enteric nervous system (ENS) in the aganglionic bowel of patients with Hirschsprung’s disease. P0-Cre/Floxed-EGFP mice in which neural crest-derived cells express EGFP were used to obtain ENS stem/progenitor cells. ENS stem/progenitor cells were transplanted into the bowel of Ret^−/− mouse, an animal model of Hirschsprung’s disease. Immunohistochemical analysis was performed to determine whether grafted cells gave rise to neurons in the recipient bowel. EGFP expressing neural crest-derived cells accounted for 7.01 ± 2.52 % of total cells of gastrointestinal tract. ENS stem/progenitor cells were isolated using flow cytometry and expanded as neurosphere-like bodies (NLBs) in a serum-free culture condition. Some cells in NLBs expressed neural crest markers, p75 and Sox10 and neural stem/progenitor cells markers, Nestin and Musashi1. Multipotency of isolated ENS stem/progenitor cells was determined as they differentiated into neurons, glial cells, and myofibloblasts in culture. When co-cultured with explants of hindgut of Ret^−/− mice, ENS stem/progenitor cells migrated into the aganglionic bowel and gave rise to neurons. ENS stem/progenitor cells used in this study appear to be clinically relevant donor cells in cell therapy to treat Hirschsprung’s disease capable of colonizing the affected bowel and giving rise to neurons.

Background: Congenital tracheal stenosis (CTS) is a rare and life-threatening disease in children. Although pulmonary artery sling (PA sling) complicated by CTS sometimes occurs, there are few reports detailing the management of CTS with PA sling. The purpose of this retrospective study was to determine the appropriate indications for surgical intervention for CTS complicated by PA sling. Methods: We evaluated 42 patients (19 males and 23 females) with the median age of 9.9±3.3 months (range, 5–34 months) with CTS complicated by PA sling who were treated at our hospital between 2005 and 2018. Twenty-eight patients received both a slide tracheoplasty and PA re-implantation, and 14 patients were managed conservatively for CTS. Among the latter, nine patients received PA re-implantation only, and five were managed conservatively without any surgery. We determined the surgical indications by retrospectively comparing the DLR value [tracheal diameter (mm)/stenotic length ratio], history of ventilator respiration, mortality rate, and post-operative course of patients at a single institution. Results: The cut-off value for the DLR was determined to be 5.9 (sensitivity: 0.929, specificity: 0.714) by using the ROC curve (AUC 0.89, P<0.05). Conclusions: A DLR value under 5.9 may serve as a new surgical indication for CTS complicated by PA sling.

Background: Hirschsprung's disease is a congenital abnormality of the enteric nervous system (ENS) presenting severe constipation soon after birth due to the lack of ganglion cells in the distal gut. Surgery for Hirschsprung's disease requires an intraoperative histopathological diagnosis to assess the extent of aganglionosis. Confocal laser endomicroscopy (CLE) is a novel endoscopic technique allowing real‐time, in vivo analysis of cellular details during ongoing endoscopy. In this study, we evaluated the possibility of a new application of CLE to provide real‐time observations of the ENS in patients with Hirschsprung's disease. In this preclinical feasibility study, we assessed the visualization of the ENS by CLE using surgically resected intestines.Methods: The subjects were nine patients who underwent pull‐through surgery for Hirschsprung's disease between September 2014 and March 2016. The colon specimens were stained with 0.1% cresyl violet and evaluated using CLE. We compared the CLE findings with those of the histopathological examination.Key Results: The ENS was clearly visualized as a ladder‐like structure in the ganglionic segment but was not observed in the aganglionic segment. Of the 69 samples, corresponding positive and negative results for both CLE and the histopathology were obtained in 61 (88%). In addition, CLE was able to visualize unique, wavy structures comprising thick nerve bundles characteristic of the aganglionic/transition zone in Hirschsprung's disease.Conclusions and Inferences: As a novel tool for visualizing the human ENS, CLE has the potential to revolutionize how pediatric surgeons identify the level of ganglionosis during surgery for Hirschsprung's disease and may be a superior alternative to intraoperative histopathological diagnosis.