Functional Lactotrophs in Induced Adenohypophysis Differentiated From Human iPS Cells

• Published in: Endocrinology (Philadelphia) 2022-03, Vol.163 (3), p.1
• Main Authors: Miyake, Natsuki, Nagai, Takashi, Suga, Hidetaka, Osuka, Satoko, Kasai, Takatoshi, Sakakibara, Mayu, Soen, Mika, Ozaki, Hajime, Miwata, Tsutomu, Asano, Tomoyoshi, Kano, Mayuko, Muraoka, Ayako, Nakanishi, Natsuki, Nakamura, Tomoko, Goto, Maki, Yasuda, Yoshinori, Kawaguchi, Yohei, Miyata, Takashi, Kobayashi, Tomoko, Sugiyama, Mariko, Onoue, Takeshi, Hagiwara, Daisuke, Iwama, Shintaro, Iwase, Akira, Inoshita, Naoko, Arima, Hiroshi, Kajiyama, Hiroaki
• Format: Article
• Language: English
• Subjects: Adrenocorticotropic hormone; Aggregates; Bromocriptine; Cell culture; Cell Culture Techniques; Cell Differentiation; Cell Line; Cell Proliferation; Cells, Cultured; Dopamine receptors; Drug discovery; Embryonic stem cells; Endocrinology; Female; Fluorescence; Fluorescence microscopy; Granule cells; Humans; Hypothalamus; Immunoelectron microscopy; Induced Pluripotent Stem Cells - drug effects; Induced Pluripotent Stem Cells - physiology; Lactation; Lactotrophs - drug effects; Lactotrophs - physiology; Nerves; Neurohormones; Neuropeptides; Peptides; Pituitary (anterior); Pituitary gland; Pituitary Gland, Anterior - cytology; Pituitary hormones; Pluripotency; Prolactin; Prolactin - biosynthesis; Prolactin-Releasing Hormone - pharmacology; Prolactin-releasing peptide; Regenerative medicine; Regulatory mechanisms (biology); Secretory vesicles; Side effects; Stem cells; Thyroid-stimulating hormone; Thyrotropin-releasing hormone; Thyrotropin-Releasing Hormone - pharmacology; Tissue engineering; Tyrosine; Tyrosine 3-monooxygenase; Vasoactive agents; Vasoactive intestinal peptide; Vasoactive Intestinal Peptide - pharmacology
• ISSN: 0013-7227; 1945-7170
• DOI: 10.1210/endocr/bqac004

Prolactin (PRL), a hormone involved in lactation, is mainly produced and secreted by the lactotrophs of the anterior pituitary (AP) gland. We previously reported a method to generate functional adrenocorticotropic hormone-producing cells by differentiating the AP and hypothalamus simultaneously from human induced pluripotent stem cells (iPSCs). However, PRL-producing cells in the induced AP have not been investigated. Here, we confirmed the presence of PRL-producing cells and evaluated their endocrine functions. We differentiated pituitary cells from human iPSCs using serum-free floating culture of embryoid-like aggregates with quick reaggregation (SFEB-q) method and evaluated the appearance and function of PRL-producing cells. Secretion of PRL from the differentiated aggregates was confirmed, which increased with further culture. Fluorescence immunostaining and immunoelectron microscopy revealed PRL-producing cells and PRL-positive secretory granules, respectively. PRL secretion was promoted by various prolactin secretagogues such as thyrotropin-releasing hormone, vasoactive intestinal peptide, and prolactin-releasing peptide, and inhibited by bromocriptine. Moreover, the presence of tyrosine hydroxylase-positive dopaminergic nerves in the hypothalamic tissue area around the center of the aggregates connecting to PRL-producing cells indicated the possibility of recapitulating PRL regulatory mechanisms through the hypothalamus. In conclusion, we generated pituitary lactotrophs from human iPSCs; these displayed similar secretory responsiveness as human pituitary cells in vivo. In the future, this is expected to be used as a model of human PRL-producing cells for various studies, such as drug discovery, prediction of side effects, and elucidation of tumorigenic mechanisms using disease-specific iPSCs. Furthermore, it may help to develop regenerative medicine for the pituitary gland.