Rat embryonic stem cell
AssigneeDainippon Sumitomo Pharma Co., Ltd.
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.
Claims34
A rat embryonic stem cell characterized by having the following properties (a)-(j):
- (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,
- (j) having an ability to produce a chimeric rat.
A rat embryonic stem cell obtained by performing a process comprising the following steps (A)-(D), under the culture conditions using a substantially serum free culture medium:
- (A) a step for dissociating an inner cell mass formed by the culture of rat blastocysts, remaining a state of cell aggregate,
- (B) a step for culturing primary embryonic stem cells resulting from the culture of the dissociated inner cell mass until it can be passaged,
- (C) a step for dissociating the primary embryonic stem cells, which have become capable of being passaged, remaining a state of cell aggregate, followed by passaging and culturing the same,
- (D) a step for further passaging and culturing the cells to establish an embryonic stem cell.
A production method of a rat embryonic stem cell which comprises performing a process comprising the following steps (A)-(D), under the culture conditions using a substantially serum free culture medium:
- (A) a step for dissociating an inner cell mass formed by the culture of rat blastocysts, remaining a state of cell aggregate,
- (B) a step for culturing primary embryonic stem cells resulting from the culture of the dissociated inner cell mass until it can be passaged,
- (C) a step for dissociating the primary embryonic stem cells, which have become capable of being passaged, remaining a state of cell aggregate, followed by passaging and culturing the same,
- (D) a step for further passaging and culturing the cells to establish an embryonic stem cell.
A subculture method of rat embryonic stem cells which comprises dissociating and passaging the cells, remaining a state of cell aggregate.
A culture medium for rat embryonic stem cell, which comprises a serum replacement reagent and rLIF.
A culture kit for rat embryonic stem cell, which comprises a serum replacement reagent and rLIF as components.
(canceled)
Dependencies are inferred from the claim text.