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US 2025/0019657 A1Published application
Generation of Adipocytes from Bovine Embryonic Stem Cell Lines
AssigneeWisconsin Alumni Research Foundation
InventorsIgor I. Slukvin, Nick Strelchenko
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Abstract
Methods for generating feeder-free and serum-free bovine embryonic stem cells (bESCs) and meat components, such as adipocytes, from bESCs are provided along with the related compositions.
Claims22
A method of culturing bovine embryonic stem cells (bESCs), the method comprising:
- culturing bESCs in a serum-free culture medium comprising an effective amount of a γ-secretase inhibitor on a matrix-coated culture vessel in feeder-free conditions;
- and
- obtaining an expanded population of bESCs expressing Oct4, Sox2, and alkaline phosphatase.
A method of producing adipocytes from bovine embryonic stem cells (bESCs), the method comprising:
- (a) culturing bESCs in a serum-free culture medium on a matrix-coated cell culture vessel in feeder-free conditions to obtain bESCs expressing Oct4, Sox2, and alkaline phosphatase, wherein the cell culture medium comprises an effective amount of γ-secretase inhibitor;
- and
- (b) dissociating the bESCs and culturing the dissociated cells on a matrix-coated cell culture vessel under serum-free medium to obtain adipocytes.
A method of producing adipocytes from bovine embryonic stem cells (bESCs), the method comprising:
- (a) culturing blastocysts on mouse embryonic fibroblasts (MEFs) in a serum-free culture medium comprising an effective amount of one or more inhibitors of the WNT pathway to obtain bESCs expressing Oct4, Sox2, and alkaline phosphatase;
- (b) culturing the bESCs from step (a) on a matrix-coated cell culture vessel in feeder-free conditions to form bESC spheres;
- and
- (c) dissociating cells from the bESC spheres and culturing the dissociated cells on a matrix-coated cell culture vessel under serum-free medium to obtain adipocytes.
1 more element
Dependencies are inferred from the claim text.