Human Minor Salivary Glands: A Readily Available Source of Salivary Stem/Progenitor Cells for Regenerative Applications.
Human Minor Salivary Glands: A Readily Available Source of Salivary Stem/Progenitor Cells for Regenerative Applications.
复制标题
人类小唾液腺：用于再生应用的唾液干细胞/祖细胞的现成来源。
DOI：
10.1007/978-1-0716-3609-1_3
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发表时间：
2024
期刊：
影响因子：
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通讯作者：
Farach-Carson,MaryC
中科院分区：
文献类型：
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作者：
Barrows,CaitlynnML;Wu,Danielle;Young,Simon;Farach-Carson,MaryC
分泌唾液腺内的常驻干细胞/祖细胞提供了潜在的治疗资源，可用于恢复慢性口干症或口干患者唾液产生所需的唾液腺再生。先前开发了从主要唾液腺（腮腺/下颌下腺）分离人类干/祖细胞（hS/PC）的方法。位于口腔和嘴唇容易接近的位置的丰富的小唾液腺可以提供额外的有价值的治疗资源。这种细胞资源的一个优点是，这些葡萄籽大小的小腺体可以通过微创外科手术从健康的捐赠者身上收获。使用小腺体的缺点是它们含有的细胞比大腺体少得多，因此收获的细胞需要在实验室中扩增以产生治疗资源。虽然早期的工作已经描述了从小唾液腺中分离增殖细胞群，可用于再生医学，但这些扩增的细胞大多数具有间充质细胞的特性，而不是分泌唾液产物的上皮细胞群的特性。在这里，我们详细描述了我们最近建立的分离和扩增从小唾液腺中分离出的 hS/PC 的方法。通过上皮祖细胞标记物 K5 和 K14 的表达对扩展的 hS/PC 群体进行上皮评估。与之前从主要唾液腺分离的可扩增细胞群一样，这些细胞也表达核 p63，这与它们在外植体培养后的扩增能力一致。当具有这些特性的 hS/PC 被封装到定制的 3D 仿生透明质酸水凝胶中时，它们将组装成保留一些祖细胞标记的微结构，同时也开始分化。分泌性粘蛋白 MUC-7 表达的增加用于证明组装的 hS/PC 微结构的分化和分泌潜力。与来自大唾液腺的 hS/PC 相比，来自小唾液腺的 hS/PC 在早期传代中往往更具异质性；因此，强烈建议使用 K5/K14/p63 作为早期质量评估工具。此外，来自小腺体的 hS/PC 对压力敏感，如果处理不当，将表现出压力反应，导致它们转变为扁平的鳞状细胞样外观，这在再生医学应用中的实用性有限。我们的结论是，正确处理的小唾液腺 hS/PC 代表了一种强大的新治疗细胞来源，可用于治疗慢性口干症患者等应用。
Resident stem/progenitor cells within the secretory salivary glands offer a potential therapeutic resource for use in the regeneration of salivary glands needed to restore saliva production in patients with chronic xerostomia, or dry mouth. Methods were developed previously to isolate human stem/progenitor cells (hS/PCs) from major salivary glands (parotid/submandibular). Abundant minor salivary glands located in readily accessible locations in the oral cavity and lip could provide an additional valuable therapeutic resource. An advantage of this cell resource is that these minor glands about the size of grape seeds can be harvested from healthy donors using minimally invasive surgical procedures. The disadvantage of using minor glands is that they contain many fewer cells than do major glands, and thus harvested cells need to be expanded in the lab to create a therapeutic resource. While earlier work has described isolation of proliferative cell populations from minor salivary glands that could be used in regenerative medicine, most of these expanded cells possess properties of mesenchymal cells rather than the epithelial population that secretes salivary products.Here, we describe in detail our recently established methods to isolate and expand hS/PCs isolated from human labial minor salivary glands. Expanded hS/PC populations are epithelial assessed by their expression of epithelial progenitor markers K5 and K14. Like expandable cell populations previously isolated from the major salivary glands, these cells also express nuclear p63, consistent with their ability to be expanded after explant culture. When hS/PCs with these properties are encapsulated into a customized 3D biomimetic hyaluronic acid-based hydrogel, they will assemble into microstructures that retain some progenitor markers while also beginning to differentiate. The increased expression of secreted mucin MUC-7 was used to demonstrate differentiation and secretory potential in assembled hS/PC microstructures. Compared to hS/PCs from major glands, those from minor salivary glands tend to be more heterogeneous in early passage; thus, use of K5/K14/p63 as an early quality assessment tool is highly recommended. Additionally, hS/PCs from minor glands are sensitive to stress and if mishandled will demonstrate a stress response that leads to their transitioning to a flat, squamous cell-like appearance that is of limited utility in regenerative medicine applications. We conclude that properly handled hS/PCs from minor salivary glands represent a powerful new source of therapeutic cells for applications including treating patients with chronic xerostomia.