Generating atrial and ventricular cardiomyocyte lineages from human pluripotent stem cells
AssigneeUniversity Health Network
InventorsGordon Keller, Stephanie Protze, Jee Hoon Lee
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Abstract
Methods are disclosed for producing populations of cardiomyocytes from pluripotent stem cells. Populations may be enriched for either atrial or ventricular cardiomyocytes and the resulting ventricular population may be essentially free of pacemaker cells. The method includes incubating pluripotent stem cells in a suitable medium with a BMP component, and an activin component, the amounts of activin may be varied to enrich for either atrial or ventricular cardiomyocytes. The enriched populations, as well as methods of using the same to treat patients in need of cardiac repair are disclosed.
Claims53
A population of cardiomyocytes enriched for ventricular cardiomyocytes, wherein said population is essentially free of pacemaker cells.
A method of producing a population of cardiomyocytes enriched for ventricular cardiomyocytes, the method comprising:
- incubating pluripotent stem cells in mesoderm induction medium, said mesoderm induction medium comprising a BMP component and an effective amount of an activin component sufficient to generate ventricular mesoderm;
- and thereafter
- culturing said incubated cells in suitable medium(s) to generate a population of cardiomyocytes enriched for ventricular cardiomyocytes.
A method of producing a population of cardiomyocytes enriched for atrial cardiomyocytes, the method comprising:
- incubating pluripotent stem cells in mesoderm induction medium, said mesoderm induction medium comprising a BMP component and an effective amount of an activin component sufficient to generate atrial mesoderm;
- and thereafter
- adding a retinoic acid component to the cells, wherein said addition of the retinoic acid component occurs during or after the incubation in mesoderm induction medium;
- and
- culturing the incubated cells so that a population of cardiomyocytes enriched for atrial cardiomyocytes is generated.
An isolated population of cardiomyocytes enriched for atrial cardiomyocytes comprising at least or about 50% of atrial cardiomyocytes, at least or about 60% of atrial cardiomyocytes, at least or about 70% of atrial cardiomyocytes, at least or about 80% of atrial cardiomyocytes, or at least or about 90% of atrial cardiomyocytes.
An isolated population of cardiomyocytes enriched for ventricular cardiomyocytes comprising at least or about 50% of ventricular cardiomyocytes, at least or about 60% of ventricular cardiomyocytes, at least or about 70% of ventricular cardiomyocytes, at least or about 80% of ventricular cardiomyocytes, or at least or about 90% of ventricular cardiomyocytes.
A process for detecting atrial mesoderm in a population of mesoderm cells, comprising detecting RALDH2, wherein a presence of RALDH2 is indicative of atrial mesoderm.
A process for detecting ventricular mesoderm in a population of mesoderm cells, comprising detecting CD235a, wherein a presence of CD235a is indicative of ventricular mesoderm.
A method for producing a population of cardiomyocytes enriched for sinoatrial nodal pacemaker cells or epicardial cells, the method comprising:
- incubating pluripotent stem cells in mesoderm induction medium, said mesoderm induction medium further comprising a BMP component and an activin component in amounts sufficient to generate ALDH+/CD235− mesoderm;
- and thereafter
- culturing said incubated cells in suitable medium(s) with one or more of WNT, FGFi and BMP to generate a population of cardiomyocytes enriched for sinoatrial nodal pacemaker cells or epicardial cells.
A method of screening or evaluating the potential cardiac toxicity of a test compound or agent, comprising the steps of exposing a population of cardiomyocytes according to any of the foregoing cell population claims to the test compound and evaluating the viability, contractility, changes in electric potentials and/or other functionalities of the cells.
Dependencies are inferred from the claim text.