Establishment of a Mouse Submandibular Salivary Gland Organ Culture
Establishment of a Mouse Submandibular Salivary Gland Organ Culture
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小鼠颌下唾液腺器官培养的建立
DOI：
10.1002/cpz1.543
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发表时间：
2022
期刊：
影响因子：
--
通讯作者：
Sakai T
中科院分区：
文献类型：
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作者：
Sakai M;Sakai T
唾液腺产生唾液，对维持口腔健康很重要。唾液保持口腔湿润，清洁口腔，帮助消化，并具有抗菌特性。唾液还有助于吞咽和说话。因此，研究唾液腺的发育在健康和疾病的背景下都是相关的。各种细胞培养方法已经被用来研究唾液腺的发育，包括二维培养细胞。在生理条件下，细胞不断与其他细胞和细胞外基质相互作用，控制细胞迁移、细胞凋亡等复杂的生物学功能，并调控基因表达。由于其中许多功能在2D培养中不能准确地表达或复制，因此使用这种培养方法的体外实验的结果通常不能反映在体内。3D培养的使用，如器官培养，帮助解决了这个问题，并已成为一种更好地反映生存生理环境中的情况的模型。在这里，我们描述了一种建立颌下腺器官培养的方案，该方案比以前的方法更简明和简单，并包括唾液腺的分离和解剖。我们还描述了使用环境应激(低氧刺激)和抑制剂(U0126、LY294002和雷帕霉素)来阐明与唾液腺发育有关的信号通路。这一方案可以为研究人员提供一种更简单、更强大的唾液腺器官培养方法，使基于器官的信号通路能够分析，以促进发育生物学研究。2022 Wiley期刊LLC基本方案1：颌下腺器官培养基本方案2：在低氧和信号通路抑制剂存在的情况下唾液腺发育的分析基本方案3：利用颌下腺器官培养进行蛋白质印迹
The salivary glands produce saliva and are important in maintaining oral health. Saliva keeps the mouth moist, cleanses the oral cavity, aids digestion, and has antibacterial properties. Saliva also helps in swallowing and speech. Investigating the development of the salivary glands is thus relevant in the context of both health and disease. Various cell culture methods have been used to study salivary gland development, including culturing cells in two dimensions (2D). Under physiological conditions, cells constantly interact with other cells and the extracellular matrix, which controls complex biological functions such as cell migration and apoptosis, and can modulate gene expression. Since many of these functions are not accurately represented or reproduced in 2D culture, the results ofin vitroexperiments using such culture methods are often not reflectedin vivo. The use of 3D cultures, such as organ cultures, has helped address this issue and has emerged as a model that better reflects thein vivophysiological environment. Here, we describe a protocol for establishing submandibular salivary gland organ culture that is more concise and simpler than previous methods and includes the separation and dissection of the salivary glands. We also describe the use of environmental stress (hypoxic stimulation) and inhibitors (U0126, LY294002, and rapamycin) to elucidate signaling pathways involved in salivary gland development. This protocol can provide researchers with a simpler and more robust method of salivary gland organ culture, enabling analysis of organ‐based signaling pathways to advance developmental biology research. © 2022 Wiley Periodicals LLC.Basic Protocol 1: Submandibular salivary gland organ cultureBasic Protocol 2: Analysis of salivary gland development in the presence of hypoxia and signaling pathway inhibitorsBasic Protocol 3: Western blotting using submandibular salivary gland organ culture