Cellular seeding and co-culture of a three dimensional fibroblast construct
Inventors
Lancaster, Jordan • Goldman, Steve
Assignees
US Department of Veterans Affairs • University of Arizona
Publication Number
US-11345894-B2
Publication Date
2022-05-31
Expiration Date
2030-04-09
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Abstract
The present invention provides methods for cellular seeding onto three-dimensional fibroblast constructs, three-dimensional fibroblast constructs seeded with muscle cells, and uses therefore.
Core Innovation
The invention provides methods for cellular seeding onto three-dimensional fibroblast constructs (3DFC), constructs comprising muscle cells adhered to a 3DFC capable of spontaneous synchronized contractions, and methods for treating disorders characterized by lack of functioning cardiomyocytes such as chronic heart failure (CHF). The methods include forcing cells in a suspension to contact and adhere to the 3DFC using forces such as centrifugation, enabling cell adhesion and growth. Additionally, the invention discloses seeding 3DFC with cell sheets, where intact confluent cells are harvested and adhered to the 3DFC, preserving cell-cell interactions.
The 3DFC comprises fibroblasts grown on a biocompatible, non-living three-dimensional framework that allows cells to form interstitial bridges and includes various types of stromal cells. Muscle cells, including cardiomyocytes or cardiac stem cells, can be seeded onto the 3DFC at specific densities and ratios to fibroblasts to generate constructs that are contractile and capable of functional electrical communication. The constructs enhance cell survival, electrical coupling via gap junctions, and synchronized contractions, providing a platform for cell-based therapies.
The problem addressed is the insufficient and disappointing results of current therapies involving injection of stem or progenitor cells directly into cardiac tissue, largely due to hostile environments lacking adequate extracellular matrix support to maintain cell survival and proper integration. The invention solves this by co-populating the 3DFC with cardiomyocytes and/or cardiac stem cells, thereby providing matrix support, improved blood flow, and angiogenic growth factors that promote engraftment, survival, and function of the seeded cells on ischemic or damaged myocardium.
Claims Coverage
The patent includes one independent claim that covers a construct comprising cells adhered to a three-dimensional fibroblast construct with specific cellular and functional features.
Construct comprising cardiomyocytes or cardiac stem cells adhered to 3DFC with functional gap junctions
The construct comprises cardiomyocytes, cardiac stem cells, or progenitors thereof adhered as at least a one-cell layer thick layer over fibroblasts on the surface of a 3DFC, with cells interconnected by functional gap junctions capable of cellular and electrical communications throughout the construct.
Method of treating disorders with constructs comprising cardiomyocytes or cardiac stem cells on 3DFC
Treatment methods involve contacting the heart of a subject suffering from a disorder characterized by a lack of functioning cardiomyocytes with an amount of the described construct effective to treat the disorder, optionally including administration of thymosin beta-4, AKT1, SDF-1, or hepatocyte growth factor.
Method for treating cardiac dysfunction with non-contracting or contracting 3DFC constructs
Methods treat impaired cardiac systolic and diastolic function by contacting the heart with the construct in an amount effective for treatment, including use of constructs non-contracting at the time of contact and optionally co-administered with therapeutic proteins.
The claims cover constructs of cardiomyocytes or cardiac stem cells adhered to a three-dimensional fibroblast construct with functional gap junctions, methods for treating cardiac disorders by applying these constructs to the heart, and associated therapeutic enhancements, providing a comprehensive coverage of the cellular construct and its medical applications.
Stated Advantages
The 3DFC co-cultured with cardiomyocytes allows cells to survive, grow, engraft, and electrically communicate through functional gap junctions, supporting spontaneous synchronized contractions.
The construct promotes angiogenesis and vascular growth by producing angiogenic factors, improving blood flow and ischemic tissue healing.
The 3DFC construct addresses poor cell retention and survival seen in direct cell injection therapies by providing matrix support and reducing cell washout.
Application of the construct results in significant improvement of cardiac function parameters such as ejection fraction, cardiac output, and end diastolic pressure in animal models of heart failure.
Documented Applications
Treatment of chronic heart failure (CHF) and related ischemic and non-ischemic cardiac disorders by applying the cell-seeded 3DFC construct onto the heart.
Promotion of healing and revascularization in ischemic heart tissue and reduction or prevention of maladaptive tissue remodeling.
Use in adjunct cardiac therapies to provide pumping assistance either through contractile integration or non-contractile functional support.
Application in treatment of stable and unstable angina, left ventricular dysfunction, and coronary artery disease by delivering constructs during procedures such as coronary artery bypass graft.
Use in combination with other cardiac treatments including pharmaceuticals, LVADs, cardiac pacemakers, cardiac wrap surgery, and enhanced external counterpulsation.
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