TY - JOUR
T1 - Generation and purification of ACTH-secreting hPSC-derived pituitary cells for effective transplantation
AU - Taga, Shiori
AU - Suga, Hidetaka
AU - Nakano, Tokushige
AU - Kuwahara, Atsushi
AU - Inoshita, Naoko
AU - Kodani, Yu
AU - Nagasaki, Hiroshi
AU - Sato, Yoshitaka
AU - Tsumura, Yusuke
AU - Sakakibara, Mayu
AU - Soen, Mika
AU - Miwata, Tsutomu
AU - Ozaki, Hajime
AU - Kano, Mayuko
AU - Watari, Kenji
AU - Ikeda, Atsushi
AU - Yamanaka, Mitsugu
AU - Takahashi, Yasuhiko
AU - Kitamoto, Sachiko
AU - Kawaguchi, Yohei
AU - Miyata, Takashi
AU - Kobayashi, Tomoko
AU - Sugiyama, Mariko
AU - Onoue, Takeshi
AU - Yasuda, Yoshinori
AU - Hagiwara, Daisuke
AU - Iwama, Shintaro
AU - Tomigahara, Yoshitaka
AU - Kimura, Toru
AU - Arima, Hiroshi
N1 - Publisher Copyright:
© 2023 The Author(s)
PY - 2023/8/8
Y1 - 2023/8/8
N2 - 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.
AB - 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.
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U2 - 10.1016/j.stemcr.2023.05.002
DO - 10.1016/j.stemcr.2023.05.002
M3 - Article
C2 - 37295423
AN - SCOPUS:85163815202
SN - 2213-6711
VL - 18
SP - 1657
EP - 1671
JO - Stem Cell Reports
JF - Stem Cell Reports
IS - 8
ER -
