Stem cell reports · 2023
Generation and purification of ACTH-secreting hPSC-derived pituitary cells for effective transplantation.
СтатьяПоддержка исследования: вне правительства СШАНазвание и аннотация: Перевод пока не подготовлен
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Taga Shiori, Suga Hidetaka, Nakano Tokushige, Kuwahara Atsushi, Inoshita Naoko, Kodani Yu, Nagasaki Hiroshi, Sato Yoshitaka, Tsumura Yusuke, Sakakibara Mayu, Soen Mika, Miwata Tsutomu, Ozaki Hajime, Kano Mayuko, Watari Kenji, Ikeda Atsushi, Yamanaka Mitsugu, Takahashi Yasuhiko, Kitamoto Sachiko, Kawaguchi Yohei, Miyata Takashi, Kobayashi Tomoko, Sugiyama Mariko, Onoue Takeshi, Yasuda Yoshinori, Hagiwara Daisuke, Iwama Shintaro, Tomigahara Yoshitaka, Kimura Toru, Arima Hiroshi
Аннотация
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.