Method for preparation of platelet from iPS cell
A method for efficiently preparing blood cells, such as mature megakaryocytes and platelets, from iPS cells is achieved in an in vitro culture system. A sac-like structure encloses hematopoietic progenitor cells, which is obtained by inoculating iPS cells onto feeder cells and then culturing the iPS cells under conditions suitable for inducing the differentiation of hematopoietic progenitor cells. Moreover, a method for producing various types of blood cells, comprises culturing hematopoietic progenitor cells enclosed in the sac-like structure under conditions suitable for inducing the differentiation of blood cells. Furthermore, a method for producing various types of blood cells, particularly megakaryocytes and platelets, is achieved without involving the sac-like structure.
1. A method for producing blood cells, the method comprising:
establishing clonal populations of human-derived induced pluripotent stem (iPS) cells;
plating the iPS cells onto OP9 cells or 10T1/2 cells, and then culturing the iPS cells under conditions suitable for inducing differentiation of hematopoietic progenitor cells, thereby forming sac-like structures containing said hematopoietic progenitor cells;
selecting human-derived iPS cell clones forming 10 or more sac-like structures per 1×10 5 cells;
plating the selected iPS cell clones onto OP9 cells or 10T1/2 cells, and then culturing under conditions suitable for inducing differentiation of hematopoietic progenitor cells, thereby forming sac-like structures containing said hematopoietic progenitor cells; wherein each of the sac-like structures takes a follicular structure and has septa composed of UEA-1 lectin positive cells;
separating cells forming the septum of the sac-like structures from the hematopoietic progenitor cells, both of which are produced by the iPS cell clones;
2. The method of claim 1 , wherein the conditions suitable for inducing the differentiation of blood cells involve the culturing of the cells for 7 to 9 days in the presence of TPO, SCF, and heparin.
3. The method of claim 1 , wherein the iPS cells are derived from introducing Oct 3/4, Sox 2, Klf4 and c-Myc into somatic cells.