High-throughput image-based chemical screening in zebrafish blastomere cell culture
AssigneeChildren's Medical Center Corporation
InventorsLeonard I. Zon, Cong Xu, Amy J. Wagers, C. Ronald Kahn
Life of this patent
Estimating the term…
Abstract
Disclosed are methods of inducing differentiation of stem into myogenic cells without gene manipulation and for inducing proliferation of satellite cells. The cells can be used as a source of cells for transplantation in a subject in need thereof. Also disclosed is a screening assay for screening test compounds using blastomere cultures.
Claims20
A method of inducing differentiation of stem cell to myogenic cell, the method comprising culturing a stem cell with at least two of:
- (i) a GSK3 pathway inhibitor;
- (ii) a compound that increases intracellular levels of 3′,5′-cyclic adenosine monophosphate (cAMP);
- and (iii) a FGF pathway activator.
-51. (canceled)
A composition comprising:
- at least two of:
- (i) a GSK3 pathway inhibitor;
- (ii) a compound that increases intracellular levels of 3′,5′-cyclic adenosine monophosphate (cAMP);
- and (iii) a FGF pathway activator.
A reagent kit for inducing differentiation from a pluripotent cell to a skeletal muscle progenitor cell, wherein the kit comprises:
- at least two of:
- (i) a GSK3 pathway inhibitor;
- (ii) a compound that increases intracellular levels of 3′,5′-cyclic adenosine monophosphate (cAMP);
- and (iii) a FGF pathway activator.
A method of increasing satellite cell proliferation, the method comprising:
- culturing a satellite cell in the presence of a compound that increases intracellular levels of 3′,5′-cyclic adenosine monophosphate.
A method of treating a subject for damaged muscle tissue or for increasing muscle mass, the method comprising co-administering to the subject in need thereof at least two of:
- (i) a GSK3 pathway inhibitor;
- (ii) a compound that increases intracellular levels of 3′,5′-cyclic adenosine monophosphate (cAMP);
- and (iii) a FGF pathway activator.
A screening assay comprising culturing a blastomere in presence of a test compound, wherein the blastomere comprises a reporter gene, wherein the reporter gene encodes a cell lineage specific marker and produces a detectable signal when expressed;
- and measuring/detecting the detectable signal, wherein a change in level or amount of the detectable signal indicating that the test compound modulates the development or function of cell lineage that express the cell lineage specific marker.
Dependencies are inferred from the claim text.