title: Mouse-isolated plexus differentiates neural crest precursors into enteric neuroblasts.
abstract: Aim of this study was to investigate, for the first time, whether isolated newborn mouse enteric plexus could induce in vitro differentiation of the vagal neural crest-derived cells into enteric neuroblasts. Fragments of the myenteric plexus were isolated from the small intestine of 6-day-old Swiss mice and were collected and stored in DMEM-F12 medium, then cultured on polymerized human fibronectin layer. The vagal portion of the neural tube, isolated from a 9.5-day-old Swiss mouse embryo, was put in the same chamber slides where the isolated myenteric plexus had been cultured for 3 days. The vagal neural crest-derived cells migrated onto the polymerized human fibronectin layer and formed a crown of cells around the neural tube. After 6 days, the cultures were stopped and studied immunohistochemically for anti-NF160 KD, anti-TH, and RetR5 antibodies to analyse the differentiation stage of the cultured cells. Analysis of results included the comparison of two culture groups: Group 1, used as control, in which vagal neural crest-derived cells were put in DMEM-F12, supplemented only with 10 % of FCS; Group 2, in which vagal neural crest-derived cells were put in the same medium as Group 1, with the addition of myenteric plexus fragments isolated from newborn mice to form the co-culture. The following results were obtained: in Group 1 the neural tubes originated a cell population strongly positive for anti-NF160 and anti-TH Ab, but negative for RetR5 Ab. This positivity was found both in the cells adjacent to the neural tube and in those migrating from it distally. The Group 2 originated cells, which after migration were positive for anti-NF160 and for anti-TH antibodies. In addition, in this culture group, the cells which migrated from the neural tube were positive for anti-RetR5 antibody. The co-culture used in this study induces the differentiation of vagal stem cells into enteric neuroblasts, cells TH+ and RetR5+. These cells, after reaching the embryonic intestine, migrate to colonize the hindgut and form the ENS. Therefore this biotechnology seems a good method to obtain in vitro enteric precursors of ENS.
authors.affiliation: Department of Paediatric Surgery, G Gaslini Children's Hospital, Genoa, Italy.
authors.forename: A
authors.initials: A
authors.lastname: Brizzolara
authors.forename: A
authors.initials: A
authors.lastname: Favre
authors.forename: K-H
authors.initials: KH
authors.lastname: Schäfer
authors.forename: A
authors.initials: A
authors.lastname: Michelazzi
authors.forename: M
authors.initials: M
authors.lastname: Sanguineti
authors.forename: G
authors.initials: G
authors.lastname: Martucciello
authors.forename: V
authors.initials: V
authors.lastname: Jasonni
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D004195:Disease Models, Animal; D017615:Enteric Nervous System / Q000166:cytology* / Q000502:physiology; D006627:Hirschsprung Disease / Q000503:physiopathology; D007150:Immunohistochemistry; D051379:Mice; D009197:Myenteric Plexus / Q000166:cytology* / Q000502:physiology; D009432:Neural Crest / Q000166:cytology* / Q000378:metabolism