R Hotta1, L S Cheng1,2, H K Graham1, W Pan1,3, N Nagy1,4, J Belkind-Gerson5,6, A M Goldstein1,6. 1. Department of Pediatric Surgery, Massachusetts General Hospital, Harvard Medical School, Boston, MA, USA. 2. Department of Surgery, University of California San Francisco, San Francisco, CA, USA. 3. Department of Pediatric Surgery, Xinhua Hospital, Shanghai Jiaotong University School of Medicine, Shanghai, China. 4. Department of Human Morphology and Developmental Biology, Faculty of Medicine, Semmelweis University, Budapest, Hungary. 5. Department of Pediatric Gastroenterology, Massachusetts General Hospital, Harvard Medical School, Boston, MA, USA. 6. Pediatric Neurogastroenterology Program, Massachusetts General Hospital, MA, USA.
Abstract
BACKGROUND: Transplanting autologous patient-derived enteric neuronal stem/progenitor cells (ENSCs) is an innovative approach to replacing missing enteric neurons in patients with Hirschsprung disease (HSCR). Using autologous cells eliminates immunologic and ethical concerns raised by other cell sources. However, whether postnatal aganglionic bowel is permissive for transplanted ENSCs and whether ENSCs from HSCR patients can be successfully isolated, cultured, and transplanted in vivo remains unknown. METHODS: ENSCs isolated from the ganglionic intestine of Ednrb(-/-) mice (HSCR-ENSCs) were characterized immunohistochemically and evaluated for their capacity to proliferate and differentiate in vitro. Fluorescently labeled ENSCs were co-cultured ex vivo with aganglionic Ednrb(-/-) colon. For in vivo transplantation, HSCR-ENSCs were labeled with lentivirus expressing green fluorescent protein (GFP) and implanted into aganglionic embryonic chick gut in ovo and postnatal aganglionic Ednrb(-/-) rectum in vivo. KEY RESULTS: HSCR-ENSCs maintain normal capacity self-renewal and neuronal differentiation. Moreover, the Ednrb(-/-) aganglionic environment is permissive to engraftment by wild-type ENSCs ex vivo and supports migratrion and neuroglial differentiation of these cells following transplantation in vivo. Lentiviral GFP-labeled HSCR-ENSCs populated embryonic chick hindgut and postnatal colon of Ednrb(-/-) HSCR, with cells populating the intermuscular layer and forming enteric neurons and glia. CONCLUSIONS & INFERENCES: ENSCs can be isolated and cultured from mice with HSCR, and transplanted into the aganglionic bowel of HSCR littermates to generate enteric neuronal networks. These results in an isogenic model establish the potential of using autologous-derived stem cells to treat HSCR and other intestinal neuropathies.
BACKGROUND: Transplanting autologous patient-derived enteric neuronal stem/progenitor cells (ENSCs) is an innovative approach to replacing missing enteric neurons in patients with Hirschsprung disease (HSCR). Using autologous cells eliminates immunologic and ethical concerns raised by other cell sources. However, whether postnatal aganglionic bowel is permissive for transplanted ENSCs and whether ENSCs from HSCR patients can be successfully isolated, cultured, and transplanted in vivo remains unknown. METHODS: ENSCs isolated from the ganglionic intestine of Ednrb(-/-) mice (HSCR-ENSCs) were characterized immunohistochemically and evaluated for their capacity to proliferate and differentiate in vitro. Fluorescently labeled ENSCs were co-cultured ex vivo with aganglionic Ednrb(-/-) colon. For in vivo transplantation, HSCR-ENSCs were labeled with lentivirus expressing green fluorescent protein (GFP) and implanted into aganglionic embryonic chick gut in ovo and postnatal aganglionic Ednrb(-/-) rectum in vivo. KEY RESULTS: HSCR-ENSCs maintain normal capacity self-renewal and neuronal differentiation. Moreover, the Ednrb(-/-) aganglionic environment is permissive to engraftment by wild-type ENSCs ex vivo and supports migratrion and neuroglial differentiation of these cells following transplantation in vivo. Lentiviral GFP-labeled HSCR-ENSCs populated embryonic chick hindgut and postnatal colon of Ednrb(-/-) HSCR, with cells populating the intermuscular layer and forming enteric neurons and glia. CONCLUSIONS & INFERENCES: ENSCs can be isolated and cultured from mice with HSCR, and transplanted into the aganglionic bowel of HSCR littermates to generate enteric neuronal networks. These results in an isogenic model establish the potential of using autologous-derived stem cells to treat HSCR and other intestinal neuropathies.
Authors: Ryo Hotta; Lincon A Stamp; Jaime P P Foong; Sophie N McConnell; Annette J Bergner; Richard B Anderson; Hideki Enomoto; Donald F Newgreen; Florian Obermayr; John B Furness; Heather M Young Journal: J Clin Invest Date: 2013-02-01 Impact factor: 14.808
Authors: Allan M Goldstein; Ryo Hotta; Lily S Cheng; Hannah K Graham; Wei Hua Pan; Nandor Nagy; Alfonso Carreon-Rodriguez Journal: J Surg Res Date: 2016-08-12 Impact factor: 2.192
Authors: Christopher R Schlieve; Kathryn L Fowler; Matthew Thornton; Sha Huang; Ibrahim Hajjali; Xiaogang Hou; Brendan Grubbs; Jason R Spence; Tracy C Grikscheit Journal: Stem Cell Reports Date: 2017-08-10 Impact factor: 7.765
Authors: Lily S Cheng; Ryo Hotta; Hannah K Graham; Jaime Belkind-Gerson; Nandor Nagy; Allan M Goldstein Journal: Pediatr Res Date: 2017-01-06 Impact factor: 3.756