Establishment of a long-term three-dimensional primary culture of mouse glandular stomach epithelial cells within the stem cell niche.
Establishment of a long-term three-dimensional primary culture of mouse glandular stomach epithelial cells within the stem cell niche.
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DOI：
10.1016/j.bbrc.2013.02.051
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发表时间：
2013-03-22
影响因子：
3.1
通讯作者：
Joh, Takashi
中科院分区：
文献类型：
--
作者：
Katano, Takahito;Ootani, Akifumi;Joh, Takashi
与小肠和结肠相比，对胃中的干细胞知之甚少，因为缺乏特异性干细胞标记物和允许长期培养的体外系统。在这里，我们描述了一个长期的三维（3D）原代胃培养系统内的干细胞龛。新生小鼠的腺胃细胞在胶原凝胶中培养3个月后产生了膨胀的球形结构。胃球壁由高度极化的上皮单层和肌成纤维细胞外层组成。上皮细胞呈高柱状，核基圆，胞浆充满粘液，表达MUC 5AC，表明分化为胃表面粘液细胞。这些细胞具有完全分化的胃表面粘液细胞的特征，如微绒毛、连接复合体、糖原和分泌颗粒。不到1%的培养上皮细胞分化为肠内分泌细胞。胃球内上皮细胞增殖活跃，并有大量凋亡细胞，表明胃球细胞更新较快。这种方法使我们能够研究在与体内环境极其相似的环境中细胞-细胞和上皮-间充质相互作用之间的信号传导的作用。(C)2013 Elsevier Inc. All rights reserved.
Compared to the small intestine and colon, little is known about stem cells in the stomach because of a lack of specific stem cell markers and an in vitro system that allows long-term culture. Here we describe a long-term three-dimensional (3D) primary gastric culture system within the stem cell niche. Glandular stomach cells from neonatal mice cultured in collagen gel yielded expanding sphere-like structures for 3 months. The wall of the gastrospheres consisted of a highly polarized epithelial monolayer with an outer lining of myofibroblasts. The epithelial cells showed a tall columnar cell shape, basal round nuclei, and mucus-filled cytoplasm as well as expression of MUC5AC, indicating differentiation into gastric surface mucous cells. These cells demonstrated the features of fully differentiated gastric surface mucous cells such as microvilli, junctional complexes, and glycogen and secretory granules. Fewer than 1% of cultured epithelial cells differentiated into enteroendocrine cells. Active proliferation of the epithelial cells and many apoptotic cells in the inner lumen revealed the rapid cell turnover in gastrospheres in vitro. This method enables us to investigate the role of signaling between cell-cell and epithelial-mesenchymal interactions in an environment that is extremely similar to the in vivo environment. (C) 2013 Elsevier Inc. All rights reserved.