Migration and differentiation of transplanted enteric neural crest-derived cells in murine model of Hirschsprung's disease.

Abstract

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.

• Authors: Ryuhei Nishikawa, Ryo Hotta, Naoki Shimojima, Shinsuke Shibata, Narihito Nagoshi, Masaya Nakamura, Yumi Matsuzaki, Hirotaka J Okano, Tatsuo Kuroda, Hideyuki Okano, Yasuhide Morikawa
• Journal: Cytotechnology (Cytotechnology) Vol. 67 Issue 4 Pg. 661-70 (Aug 2015) ISSN: 0920-9069 [Print] United States
• PMID: 25230796 (Publication Type: Journal Article)