The present invention provides a method for efficiently separating and purifying functional pituitary hormone-producing cells and/or progenitor cells thereof from differentiated tissues derived from pluripotent stem cells.

The present invention provides a method of obtaining aggregates containing a rostral hypothalamus tissue and a rostral head ectodermal tissue, a hypophysis precursor tissue and a hypophysis hormone producing cell, by using a serum-free medium (preferably substantially free of growth factor and insulins), forming homogeneous aggregates of stem cells from pluripotent stem cells such as ES cell and the like, which are plated at a high cell concentration, and subjecting the formed aggregates to floating-culture.

The present invention provides a method of obtaining aggregates containing a rostral hypothalamus tissue and a rostral head ectodermal tissue, a hypophysis precursor tissue and a hypophysis hormone producing cell, by using a serum-free medium (preferably substantially free of growth factor and insulins), forming homogeneous aggregates of stem cells from pluripotent stem cells such as ES cell and the like, which are plated at a high cell concentration, and subjecting the formed aggregates to floating-culture.

The present invention aims to provide a method for efficiently producing a cell mass containing pituitary tissue from pluripotent stem cells. A method for producing a cell mass containing pituitary tissue, including the following steps (1) and (2):

(1) a first step of suspension-culturing pluripotent stem cells to form a cell aggregate in the presence of a Wnt signal transduction pathway inhibiting substance,
(2) a second step of suspension-culturing the aggregate obtained in the first step in the presence of a BMP signal transduction pathway activating substance and a Sonic hedgehog signal transduction pathway activating substance, thereby obtaining a cell mass comprising pituitary tissue.

The presently disclosed subject matter provides for in vitro methods of inducing differentiation of stem cells (e.g., human stem cells) into somatotrophs, and differentiated cells generated by such methods. The presently disclosed subject matter also provides for uses of such cells for treating growth hormone deficiency.

The present invention provides a method of obtaining aggregates containing a rostral hypothalamus tissue and a rostral head ectodermal tissue, a hypophysis precursor tissue and a hypophysis hormone producing cell, by using a serum-free medium (preferably substantially free of growth factor and insulins), forming homogeneous aggregates of stem cells from pluripotent stem cells such as ES cell and the like, which are plated at a high cell concentration, and subjecting the formed aggregates to floating-culture.

There is provided a method for efficient differentiation induction from human pluripotent stem cells into hypothalamic neurons. Also, provided is a method for constructing, from human pluripotent stem cells, a cellular structure in which hypothalamic tissue and pituitary tissue are integrated. A cellular structure including hypothalamic tissue is obtained by a method including the steps of: culturing an aggregate of human pluripotent stem cells in suspension in a medium containing a low concentration of a bone morphogenetic protein signal transduction pathway activating substance and a low concentration of a substance acting on the Shh signaling pathway; and further culturing the cell aggregate obtained in the step in suspension in a medium containing a low concentration of a substance acting on the Shh signaling pathway.

The present invention aims to provide a method for efficiently producing a cell population containing pituitary tissue from pluripotent stem cells. The method for producing a cell population containing pituitary tissue includes the following steps (1) and (2):

(1) a first step of culturing the pluripotent stem cells in the presence of a c-jun N-terminal kinase (JNK) signal transduction pathway inhibiting substance and a Wnt signal transduction pathway inhibiting substance,
(2) a second step of culturing the cell population obtained in the first step in the presence of a BMP signal transduction pathway activating substance and a Sonic hedgehog signal transduction pathway activating substance, thereby obtaining a cell population containing pituitary tissue.

The present invention provides a method for efficiently producing a cell aggregate containing pituitary hormone-producing cells from pluripotent stem cells.

The present invention provides a method for separating pituitary hormone-producing cells and/or progenitor cells thereof, said method comprising a step for separating CD49c-expressing cells from a cell aggregate including adenohypophysis and/or a progenitor tissue thereof, and a hypothalamic neuroepithelial tissue.