Martucciello G, Brizzolara A, Favre A, Lombardi L, Bocciardi R, Sanguineti M, Pini Prato A, Jasonni V
Scientific Institute IRCCS Policlinico San Matteo, Pavia, Italy.
Eur J Pediatr Surg. 2007 Feb;17(1):34-40. doi: 10.1055/s-2007-964952.
Neural crest (NC) cells differentiate IN VITRO into neuroblasts, precursors of the enteric nervous system (ENS), when stimulated by specific agents. We developed a study aimed at establishing whether NC-derived neuroblasts can survive and colonise IN VIVO when injected into a recipient mouse gut.
The neuroblast precursors of the ENS were obtained from the vagal portion of the neural tubes of 296 CD-1 and GTROSA26 mouse embryos. The embryonic cells of GTROSA26 mice are identifiable through beta-galactosidase activity which allows recognition by blue staining. The host used in this study was the DOM/+ mouse, an animal model for Hirschsprung's disease (aganglionic megacolon). DOM/+ mouse pups (n = 43) received NC-derived cells inoculated into the seromuscular layer of the gut (33/43) or directly into the peritoneal abdominal cavity (10/43).
All DOM/+ mice survived the procedure and were sacrificed after 7 or 14 days. Histochemical staining detected implanted cells in all mice. These showed specific myenteric colonisation into the aganglionic and ganglionic gut.
The striking result of this study was the specific tropism of the injected NC-derived cells to target sites under the action of unknown chemotactic agents. This experimental procedure might represent a possible treatment option for specific forms of human ENS anomaly such as total intestinal aganglionosis.
神经嵴(NC)细胞在特定因子刺激下可在体外分化为神经母细胞,即肠神经系统(ENS)的前体。我们开展了一项研究,旨在确定注射到受体小鼠肠道内的源自NC的神经母细胞在体内能否存活并定植。
ENS的神经母细胞前体取自296只CD-1和GTROSA26小鼠胚胎神经管的迷走部分。GTROSA26小鼠的胚胎细胞可通过β-半乳糖苷酶活性识别,该活性可通过蓝色染色进行鉴定。本研究中使用的宿主是DOM/+小鼠,一种先天性巨结肠(无神经节性巨结肠)的动物模型。DOM/+小鼠幼崽(n = 43)接受了接种到肠道浆膜肌层(33/43)或直接接种到腹膜腔(10/43)的源自NC的细胞。
所有DOM/+小鼠均在手术后存活,并在7天或14天后处死。组织化学染色在所有小鼠中均检测到植入细胞。这些细胞在无神经节和有神经节的肠道中均显示出特定的肌间神经丛定植。
本研究的显著结果是,在未知趋化因子的作用下,注射的源自NC的细胞对靶位点具有特定的嗜性。这种实验方法可能代表了一种针对特定形式的人类ENS异常(如全肠无神经节症)的可能治疗选择。