# US 11,299,714 B2 · Engineered adult-like human heart tissue > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 11,299,714 B2 - Document ID: US11299714B2 - Status: Granted - Kind: B2 - Application number: 15/151,751 - Filed: 2016-05-11 - Published: 2017-01-05 - Granted: 2022-04-12 - Earliest claimed: 2015-05-11 - Assignees: The Trustees of Columbia University in the City of New York - Applicants: The Trustees of Columbia University In the City of New York - Inventors: Gordana Vunjak-Novakovic; Keith Yeager; Kacey Ronaldson; Stephen Ma; Timothy Chen - Web page: https://patentgrep.com/patent/US11299714B2 ## CPC - C12N 5/0697: {Artificial constructs associating cells of different lineages, e.g. tissue equivalents (blood vessels C12N5/0691)} - A61K 35/34: Muscles; Smooth muscle cells; Heart; Cardiac stem cells; Myoblasts; Myocytes; Cardiomyocytes (vascular smooth muscle A61K35/44) - A61K 35/545: Embryonic stem cells; Pluripotent stem cells; Induced pluripotent stem cells; Uncharacterised stem cells - A61L 27/3826: {Muscle cells, e.g. smooth muscle cells} - A61L 27/3895: {using specific culture conditions, e.g. stimulating differentiation of stem cells, pulsatile flow conditions} - C12N 5/0657: {Cardiomyocytes; Heart cells} - A61L 2430/20: for reconstruction of the heart, e.g. heart valves - C12N 2503/04: Screening or testing on artificial tissues - C12N 2506/45: from artificially induced pluripotent stem cells - C12N 2513/00: 3D culture - C12N 2529/00: Culture process characterised by the use of electromagnetic stimulation - C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00) - A61K: PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES (devices or methods specially adapted for bringing pharmaceutical products into particular physical or administering forms A61J3/00; chemical aspects of, or use of materials for deodorisation of air, for disinfection or sterilisation, or for bandages, dressings, absorbent pads or surgical articles A61L; soap compositions C11D) - A61L: METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES (preservation of bodies or disinfecting characterised by the agents employed A01N; preserving, e.g. sterilising, food or foodstuffs A23; preparations for medical, dental or toiletry purposes A61K) ## Abstract A cardiac organoid containing 3-D matter of adult human heart tissue. ## Claims ### Claim 1 An engineered three dimensional micro-tissue comprising a cell-hydrogel construct comprising electromechanically conditioned mature cardiomyocytes and dermal fibroblasts encapsulated in a fibrin hydrogel, and wherein the electromechanical conditioning comprises subjecting said construct to electrical stimulation that increases in intensity over a time period, wherein said electrical stimulation results in the micro-tissue comprising sarcomeres and a network of T-tubules. ### Claim 2 (depends on 1) The engineered three dimensional micro-tissue of claim 1, wherein the micro-tissue further comprises mitochondria or sarcoplasmic reticulum. ### Claim 3 (depends on 1) The engineered three dimensional micro-tissue of claim 1, wherein the micro-tissue exhibits a positive force-frequency relationship. ### Claim 4 (depends on 3) The engineered three dimensional micro-tissue of claim 3, wherein the positive force-frequency relationship comprises a force of about 1 to about 2 mN/mm2 at a frequency of about 0 to 6 Hz. ### Claim 5 (depends on 1) The engineered three dimensional micro-tissue of claim 1, wherein the micro-tissue exhibits a physiological response to beta-adrenergic stimulation. ### Claim 6 (depends on 5) The engineered three dimensional micro-tissue of claim 5, wherein the micro-tissue is useful for predictive drug screening, and further wherein the micro-tissue is capable of exhibiting a drug response. ### Claim 7 (depends on 1) The engineered three dimensional micro-tissue of claim 1, wherein the micro-tissue comprises sarcomeres, a network of T-tubules, mitochondria and sarcoplasmic reticulum. ### Claim 8 (depends on 1) The engineered three dimensional micro-tissue of claim 1, wherein the electromechanical stimulation comprises a frequency that increases gradually from 2 Hz at the end of week 1 to 6 Hz over a 2-week time period, and at 6 Hz for one more week. ### Claim 9 (depends on 1) The engineered three dimensional micro-tissue of claim 1, wherein the electromechanical stimulation comprises subjecting the engineered tissue to tension. ### Claim 10 (depends on 1) The engineered three dimensional micro-tissue of claim 1, wherein the sarcomeres are registers of sarcomeres having A- and I bands, and M- and Z-lines. ### Claim 11 (depends on 1) The engineered three dimensional micro-tissue of claim 1, wherein the network of T-tubules are disposed in both the longitudinal and axial orientations. ### Claim 12 An engineered three dimensional micro-tissue, comprising: a cell-hydrogel construct containing electromechanically conditioned dermal fibroblasts and mature cardiomyocytes, and wherein the electromechanical conditioning comprises subjecting said construct to electrical stimulation that increases in intensity over a time period, wherein said electrical stimulation results in the micro-tissue comprising a network of transverse-tubules, and further wherein the micro-tissue exhibits a positive force-frequency response. ### Claim 13 (depends on 12) The engineered three dimensional micro-tissue of claim 12, wherein the electromechanically conditioned micro-tissue comprises a sarcoplasmic reticulum and the micro-tissue exhibits calcium homeostasis. ### Claim 14 (depends on 12) The engineered three dimensional micro-tissue of claim 12, wherein the network of transverse tubules are disposed in both the longitudinal and axial orientations. ### Claim 15 (depends on 12) The engineered three dimensional micro-tissue of claim 12, wherein the micro-tissue exhibits expression of calcium-induced calcium release modulators. ### Claim 16 (depends on 15) The engineered three dimensional micro-tissue of claim 15, wherein the calcium release modulators are selected from the group consisting of: ryranodine receptor 2, junctophillin-2 or a combination thereof. ### Claim 17 An engineered three dimensional micro-tissue, comprising: a cell-hydrogel construct comprising electromechanically conditioned dermal fibroblasts and mature cardiomyocytes, wherein the electromechanically conditioning comprises stretching the construct between first and second pillars and subjecting said construct to electrical stimulation, said electrical stimulation including exposure to a frequency that gradually increase from 2 Hz to 6 Hz over a time period, and wherein said electrical stimulation results in the micro-tissue comprising sarcomeres and a network of transverse-tubules. ### Claim 18 (depends on 17) The engineered three-dimensional micro-tissue of claim 17, wherein the micro-tissue exhibits a positive force-frequency response. ### Claim 19 (depends on 17) The engineered three-dimensional tissue of claim 17, wherein the micro-tissue further comprises mitochondria or a sarcoplasmic reticulum. ### Claim 20 (depends on 17) The engineered three-dimensional tissue of claim 17, wherein the tissue exhibits a positive force-frequency relationship. ### Claim 21 (depends on 20) The engineered three-dimensional tissue of claim 20, wherein the positive force-frequency relationship comprises a force of about 1 to about 2 mN/mm2 at a frequency of about 0 to 6 Hz. ### Claim 22 (depends on 17) The engineered three-dimensional tissue of claim 17, wherein the micro-tissue exhibits a physiological response to beta-adrenergic stimulation. ### Claim 23 (depends on 22) The engineered three-dimensional tissue of claim 22, wherein the micro-tissue is useful for predictive drug screening, and further wherein the micro-tissue is capable of exhibiting a drug response. ### Claim 24 (depends on 17) The engineered three-dimensional micro-tissue of claim 17, wherein the mature cardiomyocytes are derived from induced pluripotent stem (iPS) cells from a patient having a disease, and further wherein the micro-tissue is used as a personalized disease model for the patient with the disease. ### Claim 25 (depends on 24) The engineered three-dimensional micro-tissue of claim 24, wherein the disease model is fora genetic disease. ### Claim 26 (depends on 25) The engineered three-dimensional micro-tissue of claim 25, wherein the genetic disease is Timothy syndrome. ### Claim 27 (depends on 24) The engineered three-dimensional micro-tissue of claim 24, wherein the disease model is for inflammation or hypoxia.