Rat embryonic stem cell
AssigneesDainippon Sumitomo Pharma Co., Ltd., National Cancer Center
InventorsTakumi Teratani, Takahiro Ochiya
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Abstract
The present invention provides a rat embryonic stem cell characterized by having the following properties of (a) expressing Oct3/4 gene and Nanog gene, (b) positive for alkaline phosphatase activity, (c) having an embryoid body forming ability, (d) expressing SSEA (Stage-Specific Embryonic Antigen)-1 and SSEA-4, (e) having the same number of chromosomes as does a normal rat cell, (f) capable of being subcultured and holding the undifferentiated state, (g) having in vitro pluripotency, (h) having a potential to differentiate for cells of three embryonic germ lineages, (i) having teratoma formation ability, and (j) having an ability to produce a chimeric rat, a method of establishing the aforementioned rat embryonic stem cell and the like.
Claims5
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).
Dependencies are inferred from the claim text.