Patent ID: 8137966
Filing Date: 2012-03-20
Classification: A01K,C12N,G01N

Abstract:
1. A method of producing a rat embryonic stem cell, which method consists essentially of the following steps (A)-(E) performed using a culture medium with 2% or less serum concentration: (A) culturing a rat blastocyst on inactivated mouse embryonic fibroblasts in a leukemia inhibitory factor (LIF)-free culture medium to form an inner cell mass in the blastocyst, (B) dissociating the inner cell mass, wherein the dissociated inner cell mass is in a cell aggregate state, (C) culturing primary embryonic stem cells resulting from a culture of the dissociated inner cell mass on inactivated mouse embryonic fibroblasts until the primary embryonic stem cells can be passaged, (D) dissociating the primary embryonic stem cells, which can be passaged, wherein the dissociated primary embryonic stem cells are in a cell aggregate state, and (E) culturing the dissociated primary embryonic stem cells on inactivated mouse embryonic fibroblasts to establish an embryonic stem cell, wherein a rat leukemia inhibitory factor (rLIF)-containing culture medium is used in steps (C)-(E).