三维培养条件下胚胎鼠唾液腺上皮细胞的自组装

背景：对唾液腺胚胎细胞在体外自组装及形态发生过程的理解,将有利于在体外构建唾液腺器官样组织的研究。目的：探讨三维培养条件下SD大鼠胚胎鼠下颌下腺上皮细胞的自组装机制。方法：将胚胎期15 d鼠的下颌下腺上皮细胞分为实验组(加入抗E-cadherin及抗β-catenin)和对照组(常规培养),在Matrigel中三维培养。结果与结论：获得的胚胎鼠下颌下腺上皮细胞均表达CK19。实验组细胞未见分枝状及腺泡样结构出现;E-cadherin 和β-catenin荧光表达弱。对照组细胞在12 d出现上述结构;胞浆中E-cadherin、β-catenin荧光表达强表达。MOD值为实验组<对照组(P < 0.01)。提示E-cadherin,β-catenin在细胞的自组装过程中有重要作用。

Self-organization and branching morphogenesis of primary salivary epithelial cells under three-dimensional culture

BACKGROUND:To understand self-assembly and branching of primary salivary epithelial cells will provide insights into reconstructing artifical salivary gland orgnoid.OBJECTIVE:To investigate the machanism of self-organization and branching mophogenesis of primary submandibular epithelial cells under three-dimensional condition.METHODS:The embryonic day 15 (E 15) mouse submandibular gland epithelium were inoculated in matrigel for 3-dimantional culture with/without (Group A/Group B) anti-E-cadherin and and anti-β-catenin. The contructs of these cells aggregation were observed under inverted microscope at days 1, 6, 12. The contructs were observed with cytokeratin 19 immunohistochemistry staining, and identified with Cadherin and β-catenin immunostaining and confocal microscopy.MAIN OUTCOME MEASURES:The constructs were observed under inverted microscope and analyzed by antibody of cytokeratin 19 with immunohistochemistry, and identified with cadherin and β-catenin immunostaining and confocal microscopy. In Group A, branching morphogenesis and acinar-like structure were not observed. In Group B, these contructs could be formed at day 12. In Group A, weak green fruorescence of E-cadherin and β-catenin expressed weakly. In Group B, they expressed strongly, and in the cytoplasm. MOD was lower in Group A than Group B (P < 0.01). E-cadherin and β-catenin are two important factors during the embryonic submandibular salivary gland epithelium self-organization.