Method for inducing trans-differentiation of cardiomyocytes based on exosome
AssigneeKorea Institute of Science and Technology
InventorsIck Chan Kwon, Sun Hwa Kim, Yoosoo Yang, Hyosuk Kim
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Abstract
The present invention relates to a method for inducing trans-differentiation of cardiomyocytes based on exosome, and more particularly, to a method for inducing trans-differentiation of a fibroblast into a cardiomyocyte, comprising the steps of: isolating exosomes in a culture medium during a process of differentiating a stem cell into the cardiomyocyte; culturing a fibroblast in a cardiomyocyte reprogramming medium containing the isolated exosomes; and culturing the fibroblast cultured in a cardiomyocyte differentiation medium containing the isolated exosomes.
Claims8
An in vitro method of inducing trans-differentiation of a fibroblast to a cardiomyocyte, the method comprising:
- a) isolating first exosomes isolated from stem cells during a mesoderm induction process and second exosomes isolated from the stem cells during a cardiac specification and maturation process;
- b) culturing a fibroblast in a cardiomyocyte reprogramming medium containing the first exosomes;
- and
- c) culturing the fibroblast in a cardiomyocyte differentiation medium containing the second exosomes isolated from the step a);
- wherein the first exosomes comprise miR-291a-3p, miR-292a-3p, miR-292a-5p, miR-293-3p, miR-293-5p, miR-295-3p, miR-294-3p, miR-292b-5p, miR-148a-3p, miR-363-3p, miR429-3p, miR291a-5p, miR-200b-3p, miR-672-5p, miR-335-3p, miR-302b-3p, miR-200a-5p, miR-881-3p, and miR-135b-5p, and the expression of miR-291a-3p, miR-292a-3p and miR-292a-5p is more than twice that of the fibroblast;
- wherein the second exosomes comprise miR-1a-3p, miR-302d-3p, miR-292a-3p, miR-291a-ep, miR-302c-3p, miR-335-3p, miR-431-5p, miR-376b-3p, miR-434-3p, miR-1b-5p, miR-434-3p, miR-293-3p, miR-291a-5p, miR-302b-3p, miR-126a-5p, miR-355-5p, miR-126b-3p, miR-148a-3p, miR-292b-3p, miR-292a-5p and miR-363-3p, and the expression of miR-1a-3p and miR-302d-3p is more than twice that of the fibroblast;
- and
- wherein the fibroblast is isolated from the same species as the stem cells.
An in vitro method for trans-differentiating a fibroblast into a cardiomyocyte, comprising:
- a) isolating exosomes in a culture medium during a process of differentiating stem cells into cardiomyocytes, wherein the exosomes are prepared by mixing first exosomes isolated from the stem cells during a mesoderm induction process and second exosomes isolated from the stem cells during a cardiac specification and maturation process;
- b) culturing the fibroblast in a cardiomyocyte reprogramming medium containing the exosomes isolated in the step a);
- and
- c) culturing the fibroblast in a cardiomyocyte differentiation medium containing the exosomes isolated in the step a) and a chemical agent for inducing cardiomyocyte differentiation;
- wherein the first exosomes comprise miR-291a-3p, miR-292a-3p, miR-292a-5p, miR-293-3p, miR-293-5p, miR-295-3p, miR-294-3p, miR-292b-5p, miR-148a-3p, miR-363-3p, miR429-3p, miR291a-5p, miR-200b-3p, miR-672-5p, miR-335-3p, miR-302b-3p, miR-200a-5p, miR-881-3p, and miR-135b-5p, and the expression of miR-291a-3p, miR-292a-3p and miR-292a-5p is more than twice that of the fibroblast;
- wherein the second exosomes comprise miR-1a-3p, miR-302d-3p, miR-292a-3p, miR-291a-ep, miR-302c-3p, miR-335-3p, miR-431-5p, miR-376b-3p, miR-434-3p, miR-1b-5p, miR-434-3p, miR-293-3p, miR-291a-5p, miR-302b-3p, miR-126a-5p, miR-355-5p, miR-126b-3p, miR-148a-3p, miR-292b-3p, miR-292a-5p and miR-363-3p, and the expression of miR-1a-3p and miR-302d-3p is more than twice that of the fibroblast;
- and
- wherein the fibroblast is isolated from the same species as the stem cells.
A composition for inducing trans-differentiation of a fibroblast into a cardiomyocyte comprising first exosomes isolated from stem cells during a mesoderm induction process, and second exosomes isolated during a cardiac specification and maturation process in the differentiation of stem cells into cardiomyocytes, and a chemical agent for inducing cardiomyocyte differentiation, wherein the chemical agent is a mixture of CHIR99021 and PD0325901;
- wherein the first exosomes comprise miR-291a-3p, miR-292a-3p, miR-292a-5p, miR-293-3p, miR-293-5p, miR-295-3p, miR-294-3p, miR-292b-5p, miR-148a-3p, miR-363-3p, miR429-3p, miR291a-5p, miR-200b-3p, miR-672-5p, miR-335-3p, miR-302b-3p, miR-200a-5p, miR-881-3p, and miR-135b-5p, and the expression of miR-291a-3p, miR-292a-3p and miR-292a-5p is more than twice that of the fibroblast;
- wherein the second exosomes comprise miR-1a-3p, miR-302d-3p, miR-292a-3p, miR-291a-ep, miR-302c-3p, miR-335-3p, miR-431-5p, miR-376b-3p, miR-434-3p, miR-1b-5p, miR-434-3p, miR-293-3p, miR-291a-5p, miR-302b-3p, miR-126a-5p, miR-355-5p, miR-126b-3p, miR-148a-3p, miR-292b-3p, miR-292a-5p and miR-363-3p, and the expression of miR-1a-3p and miR-302d-3p is more than twice that of the fibroblast;
- wherein the fibroblast is isolated from the same species as the stem cells;
- and
- wherein a mixing ratio of the first exosomes and the second exosomes is from 1:19 to 19:1.
Dependencies are inferred from the claim text.