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-9051550-B2
Publication Date
2015-06-09
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 (3DFCs), constructs comprising muscle cells adhered to these 3DFCs capable of spontaneous synchronized contractions, and therapeutic uses thereof. Specifically, the invention details how muscle cells, including cardiomyocytes or cardiac stem cells, can be seeded onto 3DFCs at particular densities and ratios with fibroblasts, forming contractile constructs that may improve heart function.
The invention addresses the problem in treating chronic heart failure (CHF) and related ischemic cardiac disorders where existing cell-based therapies, such as direct injection of stem or progenitor cells, have yielded disappointing clinical results. These failures are partly due to poor survival and engraftment of injected cells in the hostile environment of the damaged heart, lacking adequate blood supply and extracellular matrix support. The invention thus offers a solution by co-populating 3DFCs with cardiomyocytes and/or cardiac stem cells, enabling these cells to adhere, survive, engage in cell-cell communication, and improve myocardial function upon implantation.
Claims Coverage
The patent contains one independent claim focused on a specific cellular construct, covering several main inventive features related to the composition and properties of the construct.
Construct composition with cardiomyocytes or cardiac progenitors adhered to three dimensional fibroblast construct
The construct comprises cardiomyocytes, cardiac stem cells, or progenitors thereof adhered as a 1-2 cell thick layer over a layer of fibroblasts on either the top surface, bottom surface, or both surfaces of a three dimensional fibroblast construct (3DFC), with the opposite surface free of these cells.
Capacity for spontaneous synchronized contractions across entire construct
The seeded cells on the 3DFC surface are capable of initiating spontaneous contractions across the full 3DFC, and the entire construct is capable of synchronized contraction.
Seeding density and cell ratio parameters
Cardiomyocytes, cardiac stem cells, or progenitors thereof are seeded at a density between 0.5×10^6 and 5×10^6 cells/cm^2, with preferred narrower ranges including 1.3×10^6 to 5×10^6 and 1.5×10^6 to 3×10^6 cells/cm^2. The ratio of these cells to fibroblasts on the 3DFC ranges from about 1:10 to about 10:1, with preferred narrower ratios such as 1:5 to 5:1.
Human origin and engineering of cells
The cardiomyocytes, cardiac stem cells, or progenitors thereof can be of human origin and may be engineered to express therapeutic genes including thymosin beta-4 (TB4), akt murine thymoma viral oncogene homolog (AKT1), stroma cell-derived factor-1 alpha (SDF-1), and hepatocyte growth factor (HGF).
The independent claim covers a biologically active construct comprising a 3DFC with a layer of cardiomyocytes or related progenitor cells adhered in specific densities and ratios, capable of spontaneous, synchronized contractions, optionally of human origin and genetically engineered for therapeutic purposes.
Stated Advantages
Enhanced adhesion and survival of seeded cells on the 3DFC due to applied forces during seeding, avoiding poor retention seen in prior methods.
Constructs enabling spontaneous synchronized contractions across the 3DFC, demonstrating functional cellular communication via connexins.
The 3DFC provides pro-angiogenic and matrix support factors facilitating engraftment, new blood vessel formation, and cell survival in ischemic heart tissue.
The method permits treatment of chronic heart failure and related cardiac ischemic conditions with improved myocardial function, as shown by increased ejection fraction and cardiac output in preclinical models.
Documented Applications
Treatment of chronic heart failure (CHF) and related cardiac disorders characterized by lack of functioning cardiomyocytes, including ischemia without heart failure, cardiomyopathy, myocardial infarction, and valve heart diseases.
Promotion of healing and revascularization of ischemic heart tissue and prevention or reduction of maladaptive remodeling in diseased myocardium.
Use as a bioactive patch on the epicardium or other heart layers to provide contractile assistance or serve as a cell delivery system for cardiomyocytes and cardiac stem cells.
Combination with conventional therapies such as pharmaceutical agents, cardiac devices (heart pumps, LVADs, cardiac pacemakers), and cardiac wrap surgery for enhanced treatment efficacy.
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