Presaturation of supercritical CO2 with water for decellularization of matrices

Patent metadata

Field	Value
Publication number	US-11060057-B2
Application number	US-201514644241-A
Country	US
Kind code	B2
Filing date	Mar 11, 2015
Priority date	Mar 11, 2014
Publication date	Jul 13, 2021
Grant date	Jul 13, 2021

Abstract

Decellularization methods for tissue are provided. The method can include: exposing a tissue to a water-saturated, supercritical CO 2 . The method can further comprise, prior to exposing the tissue to the water-saturated, supercritical CO 2 , saturating a stream of supercritical CO 2 . The tissue can be exposed to the water-saturated, supercritical CO 2 at a treatment temperature of about 35° C. to about 40° C. (e.g., about 37° C.). In one embodiment, the water-saturated, supercritical CO 2 is completely saturated with water at the treatment temperature. The tissue can be exposed to the water-saturated, supercritical CO 2 at a constant flow rate, such as less than 3 mL/min (e.g., about 0.5 mL/min to about 2.5 mL/min).

First claim

What is claimed:

1. A decellularization method for tissue, the method comprising:
passing a stream of supercritical CO 2 through water retained in a first chamber at a flow rate of less than 3 mL/min and thereby forming a stream of water-saturated, supercritical CO 2 that is completely saturated with water;
feeding the stream of water-saturated, supercritical CO 2 that is completely saturated with water into a treatment chamber, the treatment chamber containing a tissue, such that the tissue is exposed to the water-saturated, supercritical CO 2 that is completely saturated with water; wherein the exposure causes decellularization of the tissue.

2. The decellularization method as in claim 1 , wherein the tissue is exposed to the water-saturated, supercritical CO 2 that is completely saturated with water at a treatment temperature of about 35° C. to about 40° C.

3. The decellularization method as in claim 2 , wherein the tissue is exposed to the water-saturated, supercritical CO 2 that is completely saturated with water at a treatment temperature of about 37° C.

4. The decellularization method as in claim 1 , wherein the tissue is exposed to the water-saturated, supercritical CO 2 at a constant flow rate.

5. The decellularization method as in claim 4 , wherein the flow rate is about 0.5 mL/min to about 2.5 mL/min.

6. The decellularization method as in claim 1 , wherein the water-saturated, supercritical CO 2 that is completely saturated with water has a water content of about 0.004 mole fraction to about 0.009 mole fraction.

7. The decellularization method as in claim 6 , wherein the water-saturated, supercritical CO 2 has a water content of about 0.005 mole fraction to about 0.0084 mole fraction.

8. The decellularization method as in claim 1 , wherein the tissue is exposed to the water-saturated, supercritical CO 2 that is completely saturated with water at a treatment pressure of 2000 psi.

9. The decellularization method as in claim 1 , wherein the tissue is exposed to the water-saturated, supercritical CO 2 that is completely saturated with water for a treatment time of 30 to 60 minutes per 0.25 to 0.1 gram of tissue.

10. The decellularization method as in claim 1 , further comprising following the decellularization, depressurizing the treatment chamber.

11. The decellularization method as in claim 10 , wherein the treatment chamber is depressurized at a depressurization rate of 50 psi/min.

12. The decellularization method as in claim 1 , wherein following decellularization, the weight of the tissue is from 95.4% to 98.1% of the weight of the tissue prior to decellularization.

13. The decellularization method as in claim 1 , wherein the tissue exhibits less than a one percent average weight loss upon the decellularization.

Assignees

• Univ South Carolina

Inventors

• Matthews Michael
• Casali Dominic M

Classifications

• A61L27/3691 Primary

  characterised by physical conditions of the treatment, e.g. applying a compressive force to the composition, pressure cycles, ultrasonic/sonication or microwave treatment, lyophilisation · CPC title

• A61L2430/40

  Preparation and treatment of biological tissue for implantation, e.g. decellularisation, cross-linking · CPC title

• C12N2521/00

  Culture process characterised by the use of hydrostatic pressure, flow or shear forces · CPC title

• C12N2509/00

  Methods for the dissociation of cells, e.g. specific use of enzymes · CPC title

• C12N5/0081 Primary

  Purging biological preparations of unwanted cells · CPC title