Self-organization and branching morphogenesis of primary salivary epithelial cells
Self-organization and branching morphogenesis of primary salivary epithelial cells
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DOI：
10.1089/ten.2006.0123
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发表时间：
2007-04-01
影响因子：
--
通讯作者：
Yamada, Kenneth M.
中科院分区：
文献类型：
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作者：
Wei, Cindy;Larsen, Melinda;Yamada, Kenneth M.
胚胎组织可以提供组织重组和再生所需机制的线索，但很少有研究利用原代胚胎组织来研究组织组装。为了测试组织碎片的再生能力，我们培养了胚胎第13天（E13）小鼠下颌下腺（SMG）上皮的碎片，发现小至四分之一芽的碎片保留了分支的能力。此外，我们发现完全解离的SMG上皮细胞自组织成经历显著分支的结构。对组织自组装机制的研究表明，抑制β（1）整联蛋白可防止细胞聚集，而抑制E-钙粘蛋白可阻碍聚集体致密化。免疫染色显示，细胞结构和表达模式的E-钙粘蛋白，β-连环蛋白，肌动蛋白在重组的聚合物镜那些在完整的腺体。将SMG间充质细胞添加到上皮细胞培养物中促进分支和形态分化。定量实时RT-PCR表明，聚集体表达分化标志物水通道蛋白-5（AQP 5）、催乳素诱导蛋白（PIP）和SMG蛋白C（SMGC）。总之，这些数据表明解离的SMG上皮细胞自组织并经历分支形态发生以形成具有完整腺体的结构特征和分化标志物特征的组织。这些发现提供了对自组装和分支的见解，这将有助于唾液腺和其他器官未来的再生策略。
Embryonic tissues may provide clues about mechanisms required for tissue reassembly and regeneration, but few studies have utilized primary embryonic tissue to study tissue assembly. To test the capacity of tissue fragments to regenerate, we cultured fragments of embryonic day 13 (E13) mouse submandibular gland (SMG) epithelium and found that fragments as small as a quarter-bud retain the ability to branch. Further, we found that completely dissociated SMG epithelial cells self-organize into structures that undergo significant branching. Investigation into the mechanisms involved in tissue self-assembly demonstrated that inhibition of beta(1) integrin prevents cell aggregation, while inhibition of E-cadherin hinders aggregate compaction. Immunostaining showed that the cellular architecture and expression patterns of E-cadherin, beta-catenin, and actin in the reassembled aggregates mirror those seen in intact glands. Adding SMG mesenchymal cells to the epithelial cell cultures facilitates branching and morphological differentiation. Quantitative real-time RT-PCR indicated that the aggregates express the differentiation markers aquaporin-5 (AQP5), prolactin-inducible protein (PIP), and SMG protein C (SMGC). Together, these data show that dissociated SMG epithelial cells self-organize and undergo branching morphogenesis to form tissues with structural features and differentiation markers characteristic of the intact gland. These findings provide insights into self-assembly and branching that will facilitate future regeneration strategies in the salivary gland and other organs.