Patent ID: 8796014
Filing Date: 2014-08-05
Classification: A61K,C12N

Abstract:
1. A method for producing tissue cells wherein the tissue cells are myocardial cells, the method comprising the steps of: (i) an iris-tissue-extirpating step of extirpating iris tissue from the eyeball of the animal; (ii) an iris-pigmented-epithelial-cell-separating step of separating iris pigmented epithelium from the iris tissue thus extirpated; (iii) dissociating the separated iris pigmented epithelium using a trypsin solution; (iv) obtaining pluripotent stem cells by selectively culturing iris pigment epithelial cells by a floated coagulated mass culturing technique, the iris pigmented epithelial cells separated by the steps (i)-(iii) being isolated from an eyeball of an animal, the floated coagulated mass culturing technique comprising culturing the isolated iris pigmented epithelial cells in a culturing media with rotation, the culturing media comprising serum free medium, an N2 supplement, and at least one factor selected from the group consisting of fibroblast growth factor (FGF), leukemia inhibitory factor (LIF), and stem cell factor (SCF); and (v) obtaining myocardial cells from the pluripotent stem cells by differentiating the pluripotent stem cells into myocardial cells by culturing the pluripotent stem cells under differentiation-inducing conditions comprising culturing the pluripotent stem cells for one to two months in a culture medium comprising fetal calf serum, avian serum, epidermal growth factor (EGF), and fibroblast growth factor 2 (FGF2).