Hypothalamic Contribution to Pituitary Functions Is Recapitulated In Vitro Using 3D-Cultured Human iPS Cells

Abstract

Highlights d Hypothalamic-pituitary functional units can be generated from 3D-cultured human iPS cells d Maturation of induced pituitary through simultaneous culture with hypothalamic neurons d ACTH secretion ability of induced pituitary is comparable with adult mice d Induced pituitary cells respond to low-glucose stimulation through the CRH-ACTH pathway SUMMARYThe pituitary is a major hormone center that secretes systemic hormones responding to hypothalamusderived-releasing hormones. Previously, we reported the independent pituitary induction and hypothalamic differentiation of human embryonic stem cells (ESCs).Here, a functional hypothalamic-pituitary unit is generated using human induced pluripotent stem (iPS) cells in vitro. The adrenocorticotropic hormone (ACTH) secretion capacity of the induced pituitary reached a comparable level to that of adult mouse pituitary because of the simultaneous maturation with hypothalamic neurons within the same aggregates. Corticotropin-releasing hormone (CRH) from the hypothalamic area regulates ACTH cells similarly to our hypothalamic-pituitary axis. Our induced hypothalamic-pituitary units respond to environmental hypoglycemic condition in vitro, which mimics a lifethreatening situation in vivo, through the CRH-ACTH pathway, and succeed in increasing ACTH secretion. Thus, we generated powerful hybrid organoids by recapitulating hypothalamic-pituitary development, showing autonomous maturation on the basis of interactions between developing tissues.

From Pituitary Stem Cell Differentiation to Regenerative Medicine

“…A major caveat in regenerative medicine is that human endocrine cells derived from iPSCs often do not fully recapitulate the functioning levels of normal tissues (46). However, using the previous approach for obtaining ACTH producing cells, Kasai et al 2020, demonstrated that these cells compared favorably with adult mice adenohypophysis cells, showing that the combined differentiation of pituitary and hypothalamic cells recapitulates the hormonal crosstalk between the two structures (47).…”

From Brain Organoids to Networking Assembloids: Implications for Neuroendocrinology and Stress Medicine

“… Suga et al (2011) and Ozone et al (2016) recapitulated pituitary development in 3D. Kasai et al (2020) generated a functional hypothalamus-pituitary unit. The culture methods used in the above studies have been shown to recapitulate effectively the hypothalamic and hypophyseal embryogenesis, and therefore could have applications in developmental neuroendocrinology.…”

“…Mature endocrine cells can be difficult to obtain, thus, the in vitro study of specific subtypes of human neuroendocrine cells has not been feasible; this could be overcome by the use of hPSCs. There have been several efforts to generate neuroendocrine tissues and organoids from PSCs ( Suga et al, 2011 ; Ozone et al, 2016 ; Ogawa et al, 2018 ; Kasai et al, 2020 ). Ogawa et al (2018) induced arginine vasopressin (AVP)-secreting neurons from hESCs.…”

Hypothalamic–pituitary organoid generation through the recapitulation of organogenesis

“…Several human iPS cell lines differentiated into pituitary cells using the same methods (Kasai et al, 2020). In these aggregates, after long-term maturation culture, inner hypothalamic tissues matured and expressed hypothalamic neuron markers such as CRH, and outer ACTH + cells expressed CRH receptors.…”