In vitro functional gut-like organ formation from mouse embryonic stem cells
In vitro functional gut-like organ formation from mouse embryonic stem cells
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DOI：
10.1634/stemcells.20-1-41
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发表时间：
2002-01-01
期刊：
影响因子：
5.2
通讯作者：
Tsunoda, Y
中科院分区：
文献类型：
--
作者：
Yamada, T;Yoshikawa, M;Tsunoda, Y
背景和目的。胚胎干细胞具有多能性，可在体外分化为多种细胞系。我们最近发现，胚胎干细胞可以产生一个功能类似肠道的单位，它与管腔形成三维圆顶状结构，并表现出机械活动，如自发收缩和蠕动。本研究的目的是研究胚胎干细胞衍生的收缩簇的电生理和形态学特性。通过细胞外记录检查脑电活动。免疫组织化学和电镜观察细胞形态和细胞成分。具有节律性收缩的神经元簇表现出规律的慢波，而具有高度协调蠕动的神经元簇表现出规律的慢波和自发的尖峰动作电位。在密集的网络结构中观察到Cajal间质细胞（ICC）标记物e-Kit的免疫反应性。神经元标志物PGP9.5的免疫反应性仅在伴有蠕动的簇中观察到。壁的地形结构由内上皮层和外平滑肌层组织。平滑肌层具有ICC网络，并受肠神经元支配。胚胎干细胞可以在体外分化为具有胃肠道生理和形态特征的功能性肠样器官。这种ES细胞衍生的肠道为体外研究胃肠道运动和肠道发育提供了强有力的工具，并具有阐明和治疗各种运动障碍的潜力。
Background and Aims. Embryonic stem (ES) cells have a pluripotent ability to differentiate into a variety of cell lineages in vitro. We have recently found that ES cells can give rise to a functional gut-like unit, which forms a three-dimensional dome-like structure with lumen and exhibits mechanical activity, such as spontaneous contraction and peristalsis. The aim of the present study was to investigate the electrophysiological and morphological properties of ES cell-derived contracting clusters.Methods. Electrical activity was examined by an extracellular recording. Morphology and cellular components were investigated by immunohistochemistry and electron microscopy.Results. Clusters with rhythmic contractions displayed electrical slow waves at a regular rhythm, and clusters with highly coordinated peristalsis showed regular slow waves and spontaneous spike action potentials. Immunoreactivity for e-Kit, a marker of interstitial cells of Cajal (ICC), was observed in dense network structures. Neuronal marker PGP9.5 immunoreactivity was observed only in clusters with peristalsis. The topographical structure of the wall was organized by an inner epithelial layer and outer smooth muscle layer. The smooth muscle layer was provided with an ICC network and innervated with enteric neurons.Conclusions. ES cells can differentiate into a functional gut-like organ in vitro that exhibits physiological and morphological properties characteristic of the gastrointestinal (GI) tract. This ES cell-derived gut provides a powerful tool for studying GI motility and gut development in vitro, and has potential for elucidating and treating a variety of motility disorders.