Functional Pituitary Tissue Formation.

The adenohypophysis, which mainly consists of anterior pituitary, plays important roles for endocrine systems by secreting several hormones indispensable for maintaining homeostasis. During early mouse development, the pituitary primordium (called Rathke's pouch) develops from oral ectoderm adjacent to ventral hypothalamus by interaction between these two tissues. By using mouse embryonic stem cells (ESCs), we recapitulated this in vivo micro-environment of the pituitary development and demonstrated that Rathke's pouch-like structures were self-formed from three-dimensional (3D) floating culture. The mouse ESC-derived Rathke's pouch-like structures subsequently differentiated into hormone-producing cells such as corticotrophs and somatotrophs. We have modified this technique for human pluripotent stem cells and recently reported that pituitary placodes can also be generated from human ESCs through a similar process. Here, we describe a protocol for human ESC culture for in vitro generation of 3D pituitary tissue.