Methods for tissue generation
AssigneeMilica Radisic
InventorsMilica Radisic, Yimu Zhao, Peter Backx, Naimeh Rafatian
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Abstract
The present disclosure provides ex vivo chamber-specific cardiac tissues, methods for generating the cardiac tissues in a bioreactor, and methods of using the cardiac tissues. Examples of cardiac tissues that can be generated include, but are not limited to, atrial tissues, ventricular tissues, and composite tissues having an atrial tissue connected to a ventricular tissue.
Claims14
A composite cardiac tissue grown in vitro, the composite cardiac tissue having a cylindrical geometry, the composite cardiac tissue comprising an atrial tissue at a first longitudinal end of the composite cardiac tissue and a ventricular tissue at an opposite second longitudinal end of the composite cardiac tissue, wherein the atrial tissue is connected to the ventricular tissue via a transition zone between the atrial tissue at the first longitudinal end of the composite cardiac tissue and the ventricular tissue at the second longitudinal end of the composite cardiac tissue, the transition zone comprising tissue with mixed atrial and ventricular properties, wherein a first action potential (AP) measurement taken at the atrial tissue has a first amplitude, a second AP measurement of taken at the ventricular tissue has a second amplitude higher than the first amplitude, and a third AP measurement taken at the transition zone has a third amplitude that is intermediate between the first and second amplitudes.
A method for producing a composite cardiac tissue in vitro, the composite cardiac having an atrial tissue and a ventricular tissue in connection with each other, the method comprising:
- (a) providing atrial cardiomyocytes at a first longitudinal end of a microwell and ventricular cardiomyocytes at a second opposing longitudinal end of the microwell;
- (b) applying an electrical stimulation at a first frequency to the atrial and ventricular cardiomyocytes for a first period of time, the first frequency being equal to or greater than a suprathreshold frequency;
- (c) increasing the frequency of the electrical stimulation at a rate of at least about 0.05 Hz/day until the frequency is at a second frequency of no more than about 6 Hz;
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- (d) maintaining the electrical stimulation at the second frequency for a second period of time, thereby producing the composite cardiac tissue having a cylindrical geometry, the composite cardiac tissue comprising the atrial tissue at a first longitudinal end of the composite cardiac tissue and the ventricular tissue at an opposite second longitudinal end of the composite cardiac tissue, the atrial tissue being connected to the ventricular tissue via a transition zone between the atrial tissue at the first longitudinal end of the composite cardiac tissue and the ventricular tissue at the second longitudinal end of the composite cardiac tissue, the transition zone comprising tissue with mixed atrial and ventricular properties, wherein a first action potential (AP) measurement taken at the atrial tissue has a first amplitude, a second AP measurement of taken at the ventricular tissue has a second amplitude higher than the first amplitude, and a third AP measurement taken at the transition zone has a third amplitude that is intermediate between the first and second amplitudes.
Dependencies are inferred from the claim text.