Pluripotent stem cell-derived brown adipocytes, pluripotent stem cell-derived cell aggregate, method for producing same, and cell therapy and medical therapy therefor
Provided are a method of producing brown adipocytes from pluripotent stem cells, a method of producing cell aggregates as an intermediate product thereof, pluripotent stem cell-derived cell aggregates and pluripotent stem cell-derived brown adipocytes produced by these methods, and cell therapy using the pluripotent stem cell-derived brown adipocytes. In the method of producing brown adipocytes from pluripotent stem cells, cell aggregates are produced from pluripotent stem cells by a method including the step (A), and brown adipocytes are prepared from the cell aggregates by a method including the step (B). The step (A) is a step of producing cell aggregates by non-adhesive culture of pluripotent stem cells in serum-free environment in the presence of a hematopoietic cytokine, and the step (B) is a step of producing brown adipocytes by adhesion culture of the cell aggregates in the presence of a hematopoietic cytokine.
1. A method of producing brown adipocytes, comprising the steps of:
(a) providing mouse or human pluripotent stem cells,
(b) culturing the pluripotent stem cells of step (a) in a serum-free non-adhesive culture in the presence of BMP4, VEGF, SCF, Flt3L, IL6 and IGF2 to form cell aggregates; and
(c) differentiating the cell aggregates of step (b) in an adhesive culture in the presence of BMP7, VEGF, SCF, Flt3L, IL6 and IGF3 to obtain brown adipocytes.
2. The method according to claim 1 , wherein the pluripotent stem cells are ES cells or iPS cells.
3. The method according to claim 2 , wherein the iPS cells are established using Sendai virus vectors encoding Oct3/4, Sox2, Klf4 and Myc.
4. The method according to claim 1 , wherein the pluripotent stem cells are human pluripotent stem cells.