Isolation of salivary gland stem cells and establishment of salivary gland regeneration method with tissue niche.

建立用于器官再生的组织工程技术是重建医学的重要而紧迫的问题，但很少有人尝试组织工程技术来再生唾液腺。我们展示了E13的再生能力。5胚胎颌下腺细胞(ESGCs)在I型胶原凝胶中体外培养，其结构与天然唾液腺相似。水通道蛋白5(Aquaporine 5，AQP5)在再生唾液腺的腺泡细胞中呈结构性表达，但无特征性定位。相反，AQP5在E18.5唾液腺的腺泡细胞中表达，密度较高，位于腺泡结构的管腔侧。在再生唾液腺中加入纤维连接蛋白后，AQP5的表达和定位发生了改变，这与自然唾液腺的表达和定位非常相似。在再生的唾液腺中，从第7天开始出现大的管状结构，第14天结构变大，数量增加。为了研究整合素在唾液腺发育中的作用，我们将功能扰乱抗体应用于整合素？用于再生唾液腺模型的亚单位。添加了抗？？整合素抗体明显抑制再生唾液腺的管腔形成和增大，并诱导腺泡细胞的区域化。相反，其他抗整合素抗体，特别是抗a5整合素抗体诱导管腔形成和扩大。综上所述，这些数据表明eSGCs能够再生唾液腺结构，整合素和细胞外基质之间的特异性相互作用对于再生唾液腺的分化和发育是必不可少的。

Establishment of tissue engineering techniques for organ regeneration is important and urgent issue for reconstructive medicine, but few studies have tried to regenerate salivary gland. We demonstrated the capability of regeneration of E13. 5 embryonic submandibular gland cells(eSGCs) in vitro in collagen type I gel, and the structure was similar to the native salivary gland. Aquaporine 5(AQP5) was constitutively expressed in acinar cells of regenerate salivary gland without any characteristic localization. In contrast, AQP5 expressed in acinar cells localized with fine dense concentration in the lumen side of the acinar structure in E18.5 salivary gland. By addition of fibronectin into regenerate salivary gland, the expression and localization of AQP5 was changed which was quite similar to that of native salivary gland. In regenerated salivary gland, large lumen like structures appeared from day7, and the structure enlarged and increased in number in day14. To investigate the participation of integrins in salivary gland development, we applied function-perturbing antibodies to integrin ? subunit to regenerate salivary gland model. Addition of anti-?? integrin antibody clearly inhibited lumen formation and enlargement in regenerate salivary gland, and induce compartmentalization of acinar cells. In contrast, other anti-integrin antibodies, especially anti-a5 integrin antibody induced lumen formation and enlargement. Together, these data show that eSGCs are able to regenerate salivary gland structure and specific interactions between integrins and extra cellular matrices are essential for regenerate salivary gland differentiation and development.