Department of Endocrinology and Diabetes, Graduate School of Medicine, Nagoya University, Nagoya, Aichi 466-8550, Japan; Regenerative & Cellular Medicine Kobe Center, Sumitomo Pharma Co., Ltd., Kobe, Hyogo 650-0047, Japan.
Department of Endocrinology and Diabetes, Graduate School of Medicine, Nagoya University, Nagoya, Aichi 466-8550, Japan.
Stem Cell Reports. 2023 Aug 8;18(8):1657-1671. doi: 10.1016/j.stemcr.2023.05.002. Epub 2023 Jun 8.
Pituitary organoids are promising graft sources for transplantation in treatment of hypopituitarism. Building on development of self-organizing culture to generate pituitary-hypothalamic organoids (PHOs) using human pluripotent stem cells (hPSCs), we established techniques to generate PHOs using feeder-free hPSCs and to purify pituitary cells. The PHOs were uniformly and reliably generated through preconditioning of undifferentiated hPSCs and modulation of Wnt and TGF-β signaling after differentiation. Cell sorting using EpCAM, a pituitary cell-surface marker, successfully purified pituitary cells, reducing off-target cell numbers. EpCAM-expressing purified pituitary cells reaggregated to form three-dimensional pituitary spheres (3D-pituitaries). These exhibited high adrenocorticotropic hormone (ACTH) secretory capacity and responded to both positive and negative regulators. When transplanted into hypopituitary mice, the 3D-pituitaries engrafted, improved ACTH levels, and responded to in vivo stimuli. This method of generating purified pituitary tissue opens new avenues of research for pituitary regenerative medicine.
垂体类器官是治疗垂体功能减退症的移植治疗有前途的移植物来源。在使用人类多能干细胞（hPSC）生成垂体下丘脑类器官（PHO）的自我组织培养的基础上，我们建立了使用无饲养层 hPSC 生成 PHO 和纯化垂体细胞的技术。通过对未分化的 hPSC 进行预处理，并在分化后调节 Wnt 和 TGF-β 信号，均匀且可靠地生成 PHO。使用 EpCAM（一种垂体细胞表面标志物）进行细胞分选，成功地纯化了垂体细胞，减少了非靶细胞数量。表达 EpCAM 的纯化垂体细胞重新聚集形成三维垂体球体（3D-垂体）。这些细胞表现出高促肾上腺皮质激素（ACTH）分泌能力，并对正调节剂和负调节剂均有反应。当移植到垂体功能减退的小鼠中时，3D-垂体植入后，ACTH 水平得到改善，并对体内刺激有反应。这种生成纯化垂体组织的方法为垂体再生医学开辟了新的研究途径。
Zotero 插件
