Bioreactor system and method of enhancing functionality of muscle cultured in vitro

Patent metadata

Field	Value
Publication number	US-9493735-B2
Application number	US-27967106-A
Country	US
Kind code	B2
Filing date	Apr 13, 2006
Priority date	Apr 15, 2005
Publication date	Nov 15, 2016
Grant date	Nov 15, 2016

Abstract

A method of producing organized skeletal muscle tissue from precursor muscle cells in vitro comprises: (a) providing precursor muscle cells on a support in a tissue media; then (b) cyclically stretching and relaxing the support at least twice along a first axis during a first time period; and then (c) optionally but preferably maintaining the support in a substantially static position during a second time period; and then (d) repeating steps (b) and (c) for a number of times sufficient to enhance the functionality of the tissue formed on the support and/or produce organized skeletal muscle tissue on the solid support from the precursor muscle cells.

First claim

That which is claimed is:

1. A method of producing organized skeletal muscle tissue from precursor muscle cells, comprising:
(a) providing a decellularized tissue support in a tissue media in a bioreactor, said support comprising mammalian precursor muscle cells; then
(b) cyclically stretching and relaxing said support at least twice along a first axis during a first time period, wherein said first time period is from 2 to 30 minutes in duration; and then
(c) maintaining said support in a substantially static position during a second time period, wherein said second time period is from 10 to 100 minutes in duration; and then
(d) repeating steps (b) and (c) for a number of times sufficient to enhance the functionality of the muscle tissue or produce organized skeletal muscle tissue on said support from said precursor muscle cells,
wherein the repeating steps (b) and (c) is carried out for a time of up to two weeks;
wherein said organized skeletal muscle tissue is characterized by enhancing cellular organization, contractile function and tissue formation when implanted in vivo.

2. The method of claim 1 , wherein said cyclically stretching and relaxing is carried out at least three times during said first time period.

3. The method of claim 1 , wherein said stretching comprises extending said support to a dimension at least 5% greater in length than said substantially static position.

4. The method of claim 1 , wherein said relaxing comprises retracting said support to a dimension not greater in length than said substantially static position.

5. The method of claim 1 , wherein said relaxing comprises retracting said support to a dimension not more than 5% lesser in length than said substantially static position.

6. The method of claim 1 , wherein said repeating of steps (b) and (c) is carried out for a time of five days to one week.

7. A method of producing organized skeletal muscle tissue from precursor muscle cells, comprising:
(a) providing a decellularized tissue support in a tissue media in a bioreactor, said support comprising mammalian precursor muscle cells; then
(b) cyclically stretching and relaxing said support at least twice along a first axis during a first time period, wherein said first time period is from 2 to 30 minutes in duration;
(c) cyclically stretching and relaxing said support at least twice along a second axis during said first time period; and then
(d) maintaining said support in a substantially static position during a second time period, wherein said second time period is from 10 to 100 minutes in duration; and then
(e) repeating steps (b) and (d) for a number of times sufficient to enhance the functionality of the muscle tissue or produce organized skeletal muscle tissue on said support from said precursor muscle cells,
wherein the repeating steps (b) and (d) is carried out for a time of up to two weeks;
wherein said organized skeletal muscle tissue is characterized by enhancing cellular organization, contractile function and tissue formation when implanted in vivo.

8. The method of claim 7 , wherein said cyclically stretching and relaxing along said first axis, along said second axis, or along both said first and second axis is carried out at least three times during said first time period.

9. The method of claim 7 , wherein said stretching along said first axis, along said second axis, or along both said first and second axis comprises extending said support to a dimension at least 25% greater in length than said substantially static position.

10. The method of claim 7 , wherein said relaxing along said first axis, along said second axis, or along both said first and second axis comprises retracting said support to a dimension not greater in length than said substantially static position.

11. The method of claim 7 , wherein said relaxing along said first axis, along said second axis, or along both said first and second axis comprises retracting said support to a dimension at least 5% lesser in length than said substantially static position.

12. The method of claim 7 , wherein said repeating of steps (b) and (d) is carried out for a time of five days to one week.

13. The method of claim 1 , wherein said decellularized tissue support comprises decellularized mammalian bladder.

14. The method of claim 1 , wherein said decellularized tissue support comprises decellularized porcine bladder.

15. The method of claim 7 , wherein said decellularized tissue support comprises decellularized mammalian bladder.

16. The method of claim 7 , wherein said decellularized tissue support comprises decellularized porcine bladder.

17. The method of claim 1 , wherein said organized skeletal muscle tissue on said decellularized tissue support is suturable.

18. The method of claim 7 , wherein said organized skeletal muscle tissue on said decellularized tissue support is suturable.

19. A method of producing an implantable skeletal muscle construct, comprising:
(a) providing a suturable support in a tissue media in a bioreactor, wherein said suturable support comprises decellularized mammalian bladder, and wherein said suturable support comprises mammalian precursor muscle cells; then
(b) cyclically stretching and relaxing said support at least five times along a first axis during a time period of from 2 to 30 minutes in duration; then
(c) maintaining said support in a substantially static position for a time period of from 10 to 100 minutes in duration; and then
(d) repeating steps (b) and (c) for a number of times sufficient to produce said implantable skeletal muscle construct on said support from said precursor muscle cells, wherein the repeating steps (b) and (c) is carried out for a time of up to two weeks;
wherein said implantable skeletal muscle construct is characterized by enhancing cellular organization, contractile function and tissue formation when implanted in vivo.

20. The method of claim 19 , wherein said repeating of steps (b) and (c) is carried out for a time of five days to one week.

21. The method of claim 19 , wherein said suturable support comprises decellularized porcine bladder.

22. The method of claim 19 , wherein said precursor muscle cells are human cells.

Assignees

• Yoo James
• Stitzel Joel
• Atala Anthony
• Christ George
• Univ Wake Forest Health Sciences

Inventors

• Yoo James
• Stitzel Joel
• Atala Anthony
• Christ George

Classifications

• A61L27/3826 Primary

  Muscle cells, e.g. smooth muscle cells · CPC title

• C12M21/08

  for producing artificial tissue or for ex-vivo cultivation of tissue (prostheses A61F2/00, grafts A61L27/00) · CPC title

• C12N5/0658

  Skeletal muscle cells, e.g. myocytes, myotubes, myoblasts · CPC title

• A61L27/3683

  subjected to a specific treatment prior to implantation, e.g. decellularising, demineralising, grinding, cellular disruption/non-collagenous protein removal, anti-calcification, crosslinking, supercritical fluid extraction, enzyme treatment · CPC title

• A61L2430/34

  for soft tissue reconstruction · CPC title