Primary Salivary Human Stem/Progenitor Cells Undergo Microenvironment-Driven Acinar-Like Differentiation in Hyaluronate Hydrogel Culture.
Primary Salivary Human Stem/Progenitor Cells Undergo Microenvironment-Driven Acinar-Like Differentiation in Hyaluronate Hydrogel Culture.
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DOI：
10.5966/sctm.2016-0083
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发表时间：
2017-01
影响因子：
6
通讯作者：
Pradhan-Bhatt S
中科院分区：
文献类型：
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作者：
Srinivasan PP;Patel VN;Liu S;Harrington DA;Hoffman MP;Jia X;Witt RL;Farach-Carson MC;Pradhan-Bhatt S
头颈部癌症的放射治疗通常对唾液腺有不良影响，导致口干症或严重口干，这会增加口腔感染。我们的目标是设计功能性三维（3D）唾液腺新组织，用于自体植入，以提供永久性缓解。存在获得自体成体祖细胞的迫切需要，因为胚胎和诱导多能干细胞的使用可能造成严重的风险，例如致畸性和免疫原性排斥。在这里，我们报告了一个可扩展的人口的初级唾液人干/祖细胞（hS/PC），可以重复和可扩展的分离和增殖组织活检。与腮腺的分化标志物（腺泡：MIST 1/BHLHA 15和AMY 1A;导管：K19和TFCP 2L 1）相比，这些细胞具有祖细胞标志物（K5、K14、MYC、ETV 4、ETV 5）的增加的表达。在悬浮液中生长的分离的hS/PC形成初级和次级球体，并且可以在长期3D水凝胶培养物中维持。当在用生物活性基底膜衍生肽修饰的定制的3D模块化透明质酸盐基水凝胶系统中生长时，祖细胞标志物的水平、增殖指数和hS/PC的活力得到增强。当在3D中以受控方式提供适当的微环境线索时，例如用β-肾上腺素能和胆碱能激动剂刺激，hS/PC分化成唾液产生所需的腺泡样谱系。我们得出结论，在没有抗原分选或克隆扩增的情况下分离的成体hS/PC的干/祖细胞潜力，以及它们在人类相容性培养系统中分化成专门的唾液细胞谱系的能力，使它们成为3D生物工程唾液腺应用的理想选择。干细胞转化医学2017;6：110-120
Radiotherapy for head and neck cancer often has undesirable effects on salivary glands that lead to xerostomia or severe dry mouth, which can increase oral infections. Our goal is to engineer functional, three‐dimensional (3D) salivary gland neotissue for autologous implantation to provide permanent relief. An immediate need exists to obtain autologous adult progenitor cells as the use of embryonic and induced pluripotent stem cells potentially pose serious risks such as teratogenicity and immunogenic rejection. Here, we report an expandable population of primary salivary human stem/progenitor cells (hS/PCs) that can be reproducibly and scalably isolated and propagated from tissue biopsies. These cells have increased expression of progenitor markers (K5, K14, MYC, ETV4, ETV5) compared with differentiation markers of the parotid gland (acinar: MIST1/BHLHA15 and AMY1A; ductal: K19 and TFCP2L1). Isolated hS/PCs grown in suspension formed primary and secondary spheres and could be maintained in long‐term 3D hydrogel culture. When grown in a customized 3D modular hyaluronate‐based hydrogel system modified with bioactive basement membrane‐derived peptides, levels of progenitor markers, indices of proliferation, and viability of hS/PCs were enhanced. When appropriate microenvironmental cues were provided in a controlled manner in 3D, such as stimulation with β‐adrenergic and cholinergic agonists, hS/PCs differentiated into an acinar‐like lineage, needed for saliva production. We conclude that the stem/progenitor potential of adult hS/PCs isolated without antigenic sorting or clonal expansion in suspension, combined with their ability to differentiate into specialized salivary cell lineages in a human‐compatible culture system, makes them ideal for use in 3D bioengineered salivary gland applications. Stem Cells Translational Medicine 2017;6:110–120