Methods of identifying small molecules for renewals, survival and migration of cardiac progenitors
View Patent ↗The present invention relates to a small molecule high-throughput screening assay consisting of detectably labeled cardiac progenitor cells. The invention also describes a method of identifying small molecules from the high-throughput assay affecting cardiogenesis and/or modulating cardiac progenitor cell development. Also described are methods of stimulating maturation of cardiac progenitor cells using a GSK-3β inhibitor.
1. A method of identifying a small molecule affecting cardiogenesis, wherein the small molecule is not Tamoxifen, comprising:
(a) contacting the small molecule with a cell fraction isolated from a transgenic non-human animal having a tamoxifen-dependent Cre-recombinase in the isl1 locus (isl1-mER-Cre-mER) and an Cre reporter with a fluorescent label,
(b) identifying increased fluorescence from the cell fraction;
(c) quantifying the expression of isl1+; and
(d) identifying an increase or decrease in the expression of isl1+,
wherein increased fluorescence from the cell fraction or increased expression of isl1+ is indicative of cardiogenesis,
thereby identifying a small molecule which regulates cardiogenesis.
2. The method of claim 1 , wherein the non-human animal is a rat or mouse.
3. The method of claim 1 , further comprising detecting expression of one or more growth factors selected from the group consisting of BMP4, BMP7, and FGF10.
4. A method of identifying a small molecule which regulates cardiogenesis, wherein the small molecule is not Tamoxifen, comprising:
(a) contacting the small molecule with a cell fraction isolated from a transgenic non-human animal having a tamoxifen-dependent Cre-recombinase in the isl1 locus (isl1-mER-Cre-mER) and an Cre reporter with a fluorescent label;
(b) identifying increased fluorescence from the cell fraction;
(c) quantifying the expression of isl1+; and
(d) identifying an increase or decrease in the expression of isl1+,
wherein increased fluorescence from the cell fraction or increased expression of isl1+ is indicative of cardiogenesis,
thereby identifying a small molecule which regulates cardiogenesis.
5. The method of claim 4 , wherein the non-human animal is a rat or mouse.
6. The method of claim 4 , further comprising detecting expression of one or more growth factors selected from the group consisting of BMP4, BMP7, and FGF10.