Morphogenesis and Mucus Production of Epithelial Tissues of Three Major Salivary Glands of Embryonic Mouse in 3D Culture
Morphogenesis and Mucus Production of Epithelial Tissues of Three Major Salivary Glands of Embryonic Mouse in 3D Culture
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DOI：
10.2108/zs160177
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发表时间：
2017-12-01
影响因子：
0.9
通讯作者：
Nogawa, Hiroyuki
中科院分区：
文献类型：
--
作者：
Nakao, Azusa;Inaba, Takumi;Nogawa, Hiroyuki
胚胎小鼠颌下上皮细胞植入Matrigel后，在表皮生长因子家族成员的刺激下，在两天内启动分枝形态发生。然而，尚不清楚在更长的培养时间内，末端芽是否会进一步分枝，以及产生唾液的细胞是否会在那里分化。在本研究中，我们从13天的胚胎中培养了三种主要的唾液上皮(颌下、舌下和腮腺)，在无间充质培养的情况下培养14天。在添加神经调节蛋白1、成纤维细胞生长因子1和胰岛素转铁蛋白-亚硒酸钠的培养液中，所有的上皮细胞继续生长和分支，形成大量的腺泡样结构。阿尔辛蓝染色检测粘液细胞显示，每个上皮细胞通过三种不同的方式分化，就像正常发育中看到的那样，尽管染色强度比正常发育弱。对淀粉酶基因的RT-PCR分析表明，培养14天后没有上皮细胞表达淀粉酶，这与只有出生后第7天正常发育的腮腺上皮表达淀粉酶的事实不一致。这些结果表明，在无间充质的培养中，细胞分化的进展程度低于体内培养。
Embryonic mouse submandibular epithelia initiate branching morphogenesis within two days when embedded in Matrigel and stimulated by members of the epidermal growth factor family. However, it is unknown whether the end buds further branch over longer culture periods, and whether saliva-producing cells differentiate there. In the present study, we cultivated three major (submandibular, sublingual and parotid) salivary epithelia from 13-day embryos for 14 days in mesenchyme-free cultures. All epithelia continued to grow and branch to form numerous acinus-like structures in medium supplemented with neuregulin 1, fibroblast growth factor 1, and insulintransferrin-sodium selenite. Alcian blue staining to detect mucous cells showed that each epithelium differentiated via three distinct modes, as seen in normal development, although the staining intensities were weaker than in normal development. RT-PCR analysis of the amylase gene showed that no epithelia expressed amylase after 14 days of culture, which is inconsistent with the fact that only parotid epithelium does so at postnatal day 7 during normal development. These results suggest that cytodifferentiation progresses to a lesser extent in mesenchyme-free cultures than in vivo.