In vitro investigation of the differentiation of enteric neural crest-derived cells following transplantation of aganglionic gut in a mouse model
In vitro investigation of the differentiation of enteric neural crest-derived cells following transplantation of aganglionic gut in a mouse model
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小鼠模型无神经节肠移植后肠神经嵴衍生细胞分化的体外研究
DOI：
10.1007/s00383-022-05105-2
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发表时间：
2022
影响因子：
1.8
通讯作者：
Yamataka Atsuyuki
中科院分区：
文献类型：
--
作者：
Fujiwara Naho;Miyahara Katsumi;Nakazawa-Tanaka Nana;Akazawa Chihiro;Yamataka Atsuyuki
目的细胞疗法是治疗先天性巨结肠病（Hirschsprung disease， HD）等肠道神经病变的一种很有前景的方法。最近的研究报道了来自干细胞（ENCCs）的肠神经元可以移植到HD结肠中。因此，我们研究了sox10 - venus +小鼠在移植到HD模型orEdnrbKO小鼠后肠内神经球的迁移和生成。方法从sox10 - venus +小鼠E13.5胎肠中分离单细胞悬液并分离。这些细胞在非贴壁条件下培养7天，生成神经球，并与分离的对照sox10 - venus−EdnrbKO小鼠肠道在E13.5上共培养。4天后，将这些细胞固定，检测神经元标记物Tuj1的表达。结果与HD组相比，移植的神经球在对照组肠内发生了丰富的神经细胞迁移和ENCCs分化。与对照组相比，HD结肠的平均长度和交叉点显著减少（p< 0.05）， HD小肠的平均长度和交叉点也显著减少（p< 0.05）。结论移植的ENCCs在HD肠道中不能正常分化。这些结果强调了受体肠道中神经元环境对肠神经系统发育的重要性。
PurposeCell therapy is a promising approach to treat enteric neuropathies such as Hirschsprung disease (HD). Recent studies have reported that enteric neurons derived from stem cells (ENCCs) can be grafted into the HD colon. Thus, we investigated the migration and generation of enteric neurospheres fromSOX10-VENUS+mice after transplantation into control orEdnrbKO mice, which are a model of HD.MethodsSingle-cell suspensions were isolated from the fetal guts ofSOX10-VENUS+mice E13.5 and dissociated. These cells were cultured for 7 days under non-adherent conditions to generate neurospheres, which were co-cultured with dissociated control orSOX10-VENUS−EdnrbKO mouse gut on E13.5. 4 days later, these cells were fixed and the expression of the neuronal marker, Tuj1, was evaluated.ResultsTransplanted neurospheres had undergone abundant neuronal migration and differentiation of ENCCs in the control gut compared with the HD gut. The average length and intersections were significantly decreased in HD colon compared with controls (p< 0.05), and a similar pattern was observed in the HD small intestine (p< 0.05).ConclusionsWe demonstrated that transplanted ENCCs did not differentiate properly in HD gut. These results highlight the importance of the neuronal environment in the recipient gut for enteric nervous system development.