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1. WO2020112621 - METHODS AND SYSTEMS FOR IN VITRO CARDIAC DISEASE MODELING

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[ EN ]

CLAIMS

What is claimed is:

1. A method for generating an in vitro cardiac tissue model, comprising:

forming an elongated tissue by disposing a plurality of cardiomyocytes within a culture plate;

culturing the tissue such that each end of the elongated tissue contacts one of a pair of attachment wires adhered to the culture plate; and

electrically stimulating the elongated tissue in culture.

2. The method of claim 1, wherein electrically stimulating the elongated tissue in culture further comprises:

applying an electric field along a long axis of the elongated tissue.

3. The method of claim 2, wherein electrically stimulating the elongated tissue in culture further comprises:

applying the electric field at an initial frequency of 2 Hz;

incrementally increasing the electric field to a peak frequency of 6 Hz; and

decreasing the electric field to a maintenance frequency of 3 Hz.

4. The method of claim 3, wherein electrically stimulating the elongated tissue in culture further comprises:

applying the electric field at the initial frequency of 2 Hz for 1 week; and

incrementally increasing the electric field to a peak frequency of 6 Hz over a period of 4 weeks.

5. The method of claim 4, wherein electrically stimulating the elongated tissue in culture further comprises:

applying the electric field at the maintenance frequency of 3 Hz for six months.

6. The method of claim 1, further comprising measuring force generated by the elongated tissue by imaging movement of the attachment wires.

7. The method of claim 6, wherein the attachment wires comprise a POMaC polymer; and wherein imaging movement of the attachment wires further comprises:

imaging movement of the attachment wires using UV light illumination and visible light detection.

8. The method of claim 1, wherein forming the elongated tissue further comprises:

disposing cardiac fibroblasts along with the cardiomyocytes within the culture plate.

9. The method of claim 1, wherein forming the elongated tissue further comprises:

disposing mesenchymal stem cells along with the cardiomyocytes within the culture plate.

10. The method of claim 1, wherein forming the elongated tissue further comprises:

disposing a hydrogel along with the cardiomyocytes within the culture plate.

11. The method of claim 1, wherein the cardiomyocytes comprise human induced pluripotent stem cells (hiPSCs).

12. The method of claim 11, wherein the hiPSCs are derived from a human subject with evidence of a cardiac disease.

13. The method of claim 12, wherein the cardiac disease is a polygenic disease.

14. The method of claim 13, further comprising:

analyzing gene expression in the elongated tissue to identify at least one gene related to the polygenic disease.

15. The method of claim 12, wherein the hiPSCs are at least one of: Affected D (no. A2637), Affected E (no. A2614), or Affected F (no. A2779).

16. The method of claim 11, wherein the hiPSCs are derived from a human subject without evidence of a cardiac disease.

17. The method of claim 16, wherein the hiPSCs are at least one of: Non- Affected A (no. A7156), Non-Affected B (no. 50000395), or Non- Affected C (no. U2474).

18. A kit for performing the method of claim 1, the kit comprising hiPSCs selected from the group consisting of: Non-Affected A (no. A7156), Non-Affected B (no. 50000395), Non-Affected C (no. U2474), Affected D (no. A2637), Affected E (no. A2614), and Affected F (no. A2779).

19. A kit for generating an in vitro cardiac tissue model, comprising:

a culture system including a culture plate and a pair of attachment wires; and

a plurality of hiPSC-derived cardiomyocytes from at least one of a human subject with evidence of a cardiac disease and a human subject without evidence of a cardiac disease.

20. The kit of claim 19, wherein the hiPSC-derived cardiomyocytes are from a human subject with evidence of a cardiac disease, the hiPSC-derived cardiomyocytes being selected from the group consisting of: Affected D (no. A2637), Affected E (no. A2614), and Affected F (no.

A2779).

21. The kit of claim 19, wherein the hiPSC-derived cardiomyocytes are from a human subject without evidence of a cardiac disease, the hiPSC-derived cardiomyocytes being selected from the group consisting of: Non-Affected A (no. A7156), Non-Affected B (no. 50000395), and Non-Affected C (no. U2474).

22. The kit of claim 19, wherein the culture plate further comprises:

a pair of electrodes associated with the culture plate to apply an electric field along a long axis of a tissue within the culture plate.

23. The kit of claim 19, wherein the attachment wires comprise a flexible polymer.

24. The kit of claim 23, wherein the flexible polymer comprises a POMaC polymer.

25. The kit of claim 19, further comprising a plurality of cardiac fibroblasts,

wherein the cardiac fibroblasts are disposed in the culture plate with the plurality of hiPSC-derived cardiomyocytes.

26. The kit of claim 25, further comprising a hydrogel,

wherein the cardiac fibroblasts and the plurality of hiPSC-derived cardiomyocytes are disposed in the culture plate along with the hydrogel.

27. A culture system for cardiac disease modeling, comprising:

a culture plate including a pair of anchor mechanisms; and

a plurality of hiPSC-derived cardiomyocytes from at least one of a human subject with evidence of a cardiac disease and a human subject without evidence of a cardiac disease.

28. The culture system of claim 27, wherein the culture plate further comprises:

a pair of electrodes associated with the culture plate to apply an electric field to the plurality of hiPSC-derived cardiomyocytes.

29. The culture system of claim 28, wherein the pair of anchor mechanisms comprises a pair of attachment wires each comprising a flexible polymer.

30. The culture system of claim 29, wherein the flexible polymer comprises a POMaC polymer.

31. The culture system of claim 27, further comprising a plurality of cardiac fibroblasts.

32. The culture system of claim 31, wherein the cardiac fibroblasts are disposed in the culture plate with the plurality of hiPSC-derived cardiomyocytes.

33. The culture system of claim 32, further comprising a hydrogel,

wherein the cardiac fibroblasts and the plurality of hiPSC-derived cardiomyocytes are disposed in the culture plate along with the hydrogel.

34. The culture system of claim 27, wherein the hiPSC-derived cardiomyocytes are from a human subject with evidence of a cardiac disease, the hiPSC-derived cardiomyocytes being selected from the group consisting of: Affected D (no. A2637), Affected E (no. A2614), and Affected F (no. A2779).

35. The culture system of claim 27, wherein the hiPSC-derived cardiomyocytes are from a human subject without evidence of a cardiac disease, the hiPSC-derived cardiomyocytes being selected from the group consisting of: Non-Affected A (no. A7156), Non-Affected B (no.

50000395), and Non- Affected C (no. U2474)

36. The culture system of claim 27, wherein the cardiac disease comprises heart failure.

37. A human induced pluripotent stem cell (hiPSC) from a human subject with evidence of a cardiac disease selected from the group consisting of: Affected D (no. A2637), Affected E (no. A2614), and Affected F (no. A2779).

38. A cardiomyocyte derived from the hiPSC of claim 37.

39. A human induced pluripotent stem cell (hiPSC) from a human subject without evidence of a cardiac disease selected from the group consisting of: Non-Affected A (no. A7156), Non-Affected B (no. 50000395), and Non- Affected C (no. T12474).

40. A cardiomyocyte derived from the hiPSC of claim 39.

41. A human induced pluripotent stem cell (hiPSC) from a human subject with evidence of a cardiac disease, the hiPSC consisting of cell line Affected D (no. A2637).

42. A cardiomyocyte derived from the hiPSC of claim 41.

43. A human induced pluripotent stem cell (hiPSC) from a human subject with evidence of a cardiac disease, the hiPSC consisting of cell line Affected E (no. A2614).

44. A cardiomyocyte derived from the hiPSC of claim 43.

45. A human induced pluripotent stem cell (hiPSC) from a human subject with evidence of a cardiac disease, the hiPSC consisting of cell line Affected F (no. A2779).

46. A cardiomyocyte derived from the hiPSC of claim 45.

47. A human induced pluripotent stem cell (hiPSC) from a human subject without evidence of a cardiac disease, the hiPSC consisting of cell line Non- Affected A (no. A7156).

48. A cardiomyocyte derived from the hiPSC of claim 47.

49. A human induced pluripotent stem cell (hiPSC) from a human subject without evidence of a cardiac disease, the hiPSC consisting of cell line Non-Affected B (no. 50000395).

50. A cardiomyocyte derived from the hiPSC of claim 49.

51. A human induced pluripotent stem cell (hiPSC) from a human subject without evidence of a cardiac disease, the hiPSC consisting of cell line Non-Affected C (no. U2474).

52. A cardiomyocyte derived from the hiPSC of claim 51.

53. A kit for screening a drug candidate comprising a cardiomyocyte of claim 38 and a cardiomyocyte of claim 40.

54. A method for screening a drug candidate comprising applying the drug candidate to the cardiomyocytes of claim 53.