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Disruption of these interactions underlies the pathogenesis of intestinal aganglionosis in Hirschsprung disease (HSCR). ENSC therapy has been proposed as a possible treatment for HSCR, but whether the survival and development of postnatal-derived ENSCs similarly rely on signals from the mesenchymal environment is unknown and has important implications for developing protocols to expand ENSCs for cell transplantation therapy. Enteric neural crest-derived cells (ENCDCs) and EMCs were cultured from the small intestine of Wnt1-Rosa26-tdTomato mice. EMCs promoted the expansion of ENCDCs 9.5-fold by inducing ENSC properties, including expression of Nes, Sox10, Sox2, and Ngfr. EMCs enhanced the neurosphere-forming ability of ENCDCs, and this persisted after withdrawal of the EMCs. These effects were mediated by paracrine factors and several ligands known to support neural stem cells were identified in EMCs. Using the optimized expansion procedures, neurospheres were generated from small intestine of the Ednrb \u2212\/\u2212 mouse model of HSCR. These ENSCs had similar proliferative and migratory capacity to Ednrb +\/+ ENSCs, albeit neurospheres contained fewer neurons. ENSCs derived from Ednrb \u2212\/\u2212 mice generated functional neurons with similar calcium responses to Ednrb +\/+ ENSCs and survived after transplantation into the aganglionic colon of Ednrb \u2212\/\u2212 recipients. EMCs act as supporting cells to ENSCs postnatally via an array of synergistically acting paracrine signaling factors. These properties can be leveraged to expand autologous ENSCs from patients with HSCR mutations for therapeutic application.<\/jats:p>","DOI":"10.1002\/stem.3388","type":"journal-article","created":{"date-parts":[[2021,11,11]],"date-time":"2021-11-11T09:59:49Z","timestamp":1636624789000},"page":"1236-1252","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":33,"title":["Enteric mesenchymal cells support the growth of postnatal enteric neural stem cells"],"prefix":"10.1093","volume":"39","author":[{"given":"Rhian","family":"Stavely","sequence":"first","affiliation":[{"name":"Department of Pediatric Surgery \u00a0Massachusetts General Hospital, Harvard Medical School, Boston, Massachusetts, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Sukhada","family":"Bhave","sequence":"additional","affiliation":[{"name":"Department of Pediatric Surgery \u00a0Massachusetts General Hospital, Harvard Medical School, Boston, Massachusetts, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Wing Lam N.","family":"Ho","sequence":"additional","affiliation":[{"name":"Department of Pediatric Surgery \u00a0Massachusetts General Hospital, Harvard Medical School, Boston, Massachusetts, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Minhal","family":"Ahmed","sequence":"additional","affiliation":[{"name":"Department of Pediatric Surgery \u00a0Massachusetts General Hospital, Harvard Medical School, Boston, Massachusetts, USA"},{"name":"College of Engineering \u00a0Northeastern University, Boston, Massachusetts, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Weikang","family":"Pan","sequence":"additional","affiliation":[{"name":"Department of Pediatric Surgery \u00a0Massachusetts General Hospital, Harvard Medical School, Boston, Massachusetts, USA"},{"name":"Department of Pediatric Surgery \u00a0The Second Affiliated Hospital of Xi'an Jiaotong University, Xi'an City, People's Republic of China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ahmed A.","family":"Rahman","sequence":"additional","affiliation":[{"name":"Department of Pediatric Surgery \u00a0Massachusetts General Hospital, Harvard Medical School, Boston, Massachusetts, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jessica","family":"Ulloa","sequence":"additional","affiliation":[{"name":"Department of Pediatric Surgery \u00a0Massachusetts General Hospital, Harvard Medical School, Boston, Massachusetts, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Nicole","family":"Bousquet","sequence":"additional","affiliation":[{"name":"Department of Pediatric Surgery \u00a0Massachusetts General Hospital, Harvard Medical School, Boston, Massachusetts, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Meredith","family":"Omer","sequence":"additional","affiliation":[{"name":"Department of Pediatric Surgery \u00a0Massachusetts General Hospital, Harvard Medical School, Boston, Massachusetts, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Richard","family":"Guyer","sequence":"additional","affiliation":[{"name":"Department of Pediatric Surgery \u00a0Massachusetts General Hospital, Harvard Medical School, Boston, Massachusetts, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Nandor","family":"Nagy","sequence":"additional","affiliation":[{"name":"Department of Pediatric Surgery \u00a0Massachusetts General Hospital, Harvard Medical School, Boston, Massachusetts, USA"},{"name":"Department of Anatomy, Histology and Embryology, Faculty of Medicine \u00a0Semmelweis University, Budapest, Hungary"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Allan M.","family":"Goldstein","sequence":"additional","affiliation":[{"name":"Department of Pediatric Surgery \u00a0Massachusetts General Hospital, Harvard Medical School, Boston, Massachusetts, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9890-8241","authenticated-orcid":false,"given":"Ryo","family":"Hotta","sequence":"additional","affiliation":[{"name":"Department of Pediatric Surgery \u00a0Massachusetts General Hospital, Harvard Medical School, Boston, Massachusetts, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"286","published-online":{"date-parts":[[2021,5,8]]},"reference":[{"issue":"12","key":"2022022511244874800_stem3388-bib-0001","doi-asserted-by":"crossref","first-page":"2273","DOI":"10.1111\/j.1440-1746.2006.04750.x","article-title":"Long-term continence after surgery for Hirschsprung's disease","volume":"22","author":"Catto-Smith","year":"2007","journal-title":"J Gastroenterol Hepatol"},{"issue":"3","key":"2022022511244874800_stem3388-bib-0002","doi-asserted-by":"crossref","first-page":"1182","DOI":"10.1172\/JCI65963","article-title":"Transplanted progenitors generate functional enteric 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