Functional Lactotrophs in Induced Adenohypophysis Differentiated From Human iPS Cells
Functional Lactotrophs in Induced Adenohypophysis Differentiated From Human iPS Cells
复制标题
DOI：
10.1210/endocr/bqac004
复制
发表时间：
2022-03-01
期刊：
影响因子：
4.8
通讯作者：
Kajiyama, Hiroaki
中科院分区：
文献类型：
--
作者：
Miyake, Natsuki;Nagai, Takashi;Kajiyama, Hiroaki
催乳素（PRL）是一种与哺乳有关的激素，主要由垂体前叶（AP）腺的催乳细胞产生和分泌。我们以前报道了一种方法，以产生功能性促肾上腺皮质激素分泌细胞分化的AP和下丘脑同时从人类诱导多能干细胞（iPSC）。然而，PRL产生细胞在诱导AP尚未调查。在这里，我们证实了PRL产生细胞的存在，并评估了它们的内分泌功能。我们使用无血清漂浮培养的胚胎样聚集体与快速再聚集（SFEB-q）方法从人iPSCs分化垂体细胞，并评估PRL产生细胞的外观和功能。PRL从分化的聚集体的分泌被证实，这增加了进一步的文化。荧光免疫染色和免疫电镜分别显示PRL产生细胞和PRL阳性分泌颗粒。PRL分泌促进各种催乳素促分泌素，如促甲状腺激素释放激素，血管活性肠肽，催乳素释放肽，溴隐亭抑制。此外，酪氨酸羟化酶阳性的多巴胺能神经的存在下丘脑组织区域周围的聚集体连接到PRL产生细胞的中心表明通过下丘脑重演PRL调节机制的可能性。总之，我们从人iPSC产生垂体催乳素细胞;这些细胞在体内显示出与人垂体细胞相似的分泌反应。在未来，这有望被用作各种研究的人类PRL产生细胞的模型，如药物发现，副作用的预测，以及使用疾病特异性iPSC阐明肿瘤发生机制。此外，它可能有助于开发脑下垂体的再生医学。
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.