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Last Updated: January 23, 2020

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Claims for Patent: 10,188,683

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Summary for Patent: 10,188,683
Title:Tissue engineering of lung
Abstract: The present invention relates to compositions comprising a decellularized tissue. The present invention also provides an engineered three dimensional lung tissue exhibiting characteristics of a natural lung tissue. The engineered tissue is useful for the study of lung developmental biology and pathology as well as drug discovery.
Inventor(s): Calle; Elizabeth (Danbury, CT), Niklason; Laura E. (Greenwich, CT), Petersen; Thomas (Setauket, NY), Gui; Liqiong (New Haven, CT)
Assignee: Yale University (New Haven, CT)
Application Number:14/625,080
Patent Claims:1. A method of making a decellularized tissue, comprising perfusing a natural tissue comprising a capillary network with a decellularization solution, wherein the natural tissue is isolated from a mammal, wherein the decellularization solution comprises a solution hypertonic to cells in the tissue, a zwitterionic detergent, and a chelating agent, and wherein the decellularization solution removes cellular material and retains collagen, capillary structure, and structural integrity of the matrix similar to the natural tissue, further comprising monitoring a perfusion pressure during the perfusing and adjusting the perfusion pressure to maintain a pressure of less than 30 mmHg.

2. The method of claim 1, wherein the natural tissue is a lung tissue comprising an airway network.

3. The method of claim 2, wherein the decellularization solution is perfused through the airway network of the lung tissue.

4. The method of claim 2, further comprising lavaging the airway network with the decellularization solution prior to the perfusing.

5. The method of claim 2, wherein the lung tissue comprises a lobe.

6. The method of claim 1, further comprising replacing the decellularization solution at least once during the perfusing.

7. The method of claim 1, further comprising monitoring a perfusion pressure during the perfusing and adjusting the perfusion pressure to maintain a pressure of less than 17 mmHg.

8. The method of claim 1, wherein the perfusing continues until substantially all cells and antigenic molecules are removed from the tissue.

9. The method of claim 1, wherein the decellularization solution further comprises a buffer.

10. The method of claim 9, wherein the buffer is phosphate-buffered saline (PBS).

11. The method of claim 1, wherein the decellularization solution further comprises an enzyme.

12. The method of claim 11, wherein the enzyme is selected from the group consisting of a collagenase, a dispase, a DNase, a protease, and combinations thereof.

13. The method of claim 1, wherein the decellularization solution further comprises an enzyme inhibitor.

14. The method of claim 13, wherein the enzyme inhibitor is selected from the group consisting of a protease inhibitor, a nuclease inhibitor, a collagenase inhibitor, and combinations thereof.

15. The method of claim 1, wherein the decellularization solution further comprises a vasodilator.

16. The method of claim 1, wherein the decellularization solution is perfused at a pressure less than about 10 mmHg.

17. The method of claim 1, wherein the zwitterionic detergent is 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate (CHAPS).

18. The method of claim 1, wherein the hypertonic solution is a hypertonic sodium chloride solution and the chelating agent is ethylenedinitrilo-tetraacetic acid (EDTA).

19. The method of claim 1, wherein the hypertonic solution is about 1M sodium chloride (NaCl), the detergent is about 8 mM CHAPS, and the chelating agent is about 25 mM EDTA.

20. A method of making a decellularized lung tissue, comprising perfusing a natural lung tissue with a decellularization solution at a pressure of 20 mmHg or less until substantially all of the cells and antigenic molecules are removed from the natural lung tissue, wherein the decellularization solution comprises a NaCl solution hypertonic to cells in the lung tissue, a zwitterionic detergent, and a chelating agent, and wherein the decellularized lung tissue comprises at least a lobe, an airway network, and a vascular network and retains collagen, capillary structure and structural integrity of the matrix similar to the natural lung tissue, and wherein the natural lung tissue is isolated from a mammal.

21. The method of claim 20, wherein the lung tissue comprises a whole lung.

22. The method of claim 20, wherein the pressure is less than 17 mmHg.

23. The method of claim 20, wherein the zwitterionic detergent is CHAPS and the chelating agent is EDTA.

24. The method of claim 20, wherein the hypertonic NaCl solution is about 1M NaC1, the zwitterionic detergent is about 8 mM CHAPS, and the chelating agent is about 25 mM EDTA.

25. The method of claim 20, wherein the perfusing removes substantially all MHC Class I and II antigens.

Details for Patent 10,188,683

Applicant Tradename Biologic Ingredient Dosage Form BLA Number Approval Date Patent No. Assignee Estimated Patent Expiration Status Orphan Source
Smith And Nephew SANTYL collagenase OINTMENT;TOPICAL 101995 001 1965-06-04   Start Trial Yale University (New Haven, CT) 2029-02-04 RX search
>Applicant >Tradename >Biologic Ingredient >Dosage Form >BLA >Number >Approval Date >Patent No. >Assignee >Estimated Patent Expiration >Status >Orphan >Source

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