US5158867A
Expired / Lapsed
US
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Method for cryopreserving blood vessels
Assignee: Medical University of South Carolina
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Filing: 1990-01-23
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Granted: 1992-10-27
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Expires: 2010-01-23
Abstract
Abstract A device for use in cryopreservation of blood vessels comprising a pair of stylets insertable into the ends of a dissected blood vessel segment. The stylets are mountable on a support track whereby the blood vessel can be distended and supported during cryopreservation procedures. Also disclosed is a freezing and thawing profile capable of maximizing endothelial cell survival. The use of chondroitin sulfate or similar compound is discussed as a novel cryoprotectant.
Inventors
- 👤 Robert T. McNally
- 👤 Cameron McCaa
- 👤 Kelvin G. M. Brockbank
- 👤 Albert E. Heacox
- 👤 Harvey L. Bank
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-04 09:14
CPC Classifications
A; A01; A01N; A01N1/00; A01N1/10; A01N1/16; A01N1/162
Claims Analysis
Total Claims: 12
Claim 1
Independent Claim
73 words
A method of maintaining intact a layer of endothelial cells present in a blood vessel during cryopreservation comprising: i) contacting said blood vessel with an effective amount of a cryoprotectant composition comprising a medium for freezing said blood vessel, a cryoprotectively effective concentration of a cell penetrating cryoprotectant and a cryoprotectively effective concentration of a glycosaminoglycan; and ii) maintaining said blood vessel in contact with said composition at a temperature below -100° C.
Claim 2
Independent Claim
73 words
A method of maintaining intact a layer of endothelial cells present in a blood vessel during cryopreservation comprising: i) contacting said blood vessel with an effective amount of a cryoprotectant composition comprising a medium for freezing said blood vessel, a cryoprotectively effective concentration of a cell penetrating cryoprotectant and a cryoprotectively effective concentration of a glycosaminoglycan; and ii) maintaining said blood vessel in contact with said composition at a temperature below -100° C.
Claim 3
Dependent Claim
13 words
The method according to claim 1 wherein said composition further comprises an antibiotic.
Claim 4
Dependent Claim
12 words
The method according to claim 1 wherein the glycosaminoglycan is chondroitin sulfate.
Claim 5
Dependent Claim
13 words
The method according to claim 1 wherein said cell penetrating cryoprotectant is dimethylsulfoxide.
Claim 6
Dependent Claim
73 words
The method of claim 1 wherein the freezing schedule comprises: (a) placing the blood vessel into a freezing chamber; (b) cooling the sample at a suitable rate until the sample reaches about -2° C.; (c) cooling the sample after substantial completion of phase change at a suitable rate; (d) removing the blood vessel from the chamber; (e) placing the blood vessel in a liquid nitrogen refrigerator at a temperature of about -196° C.
Claim 7
Dependent Claim
18 words
The method of claim 5 wherein the cooling rate is in a range of from about 0.1°-100° C./min.
Claim 8
Dependent Claim
18 words
The method of claim 6 wherein the cooling rate is in a range of from about 0.1°-30° C./min.
Claim 9
Dependent Claim
18 words
The method of claim 7 wherein the cooling rate is in a range of from about 0.3°-1° C./min.
Claim 10
Dependent Claim
18 words
The method of claim 8 wherein the cooling rate is in a range of from about 0.5°-1° C./min.
Claim 11
Dependent Claim
18 words
The method of claim 9 wherein the cooling rate is in a range of from about 0.5° C./min.
Claim 12
Dependent Claim
247 words
The method of claim 5 wherein the freezing schedule comprises: (a) placing the blood vessel into a freezing chamber, then (b) cooling the chamber from an initial temperature of about -10° C. at a rate of about 0.01° C. per min. until the sample reaches about +4° C.; then (c) cooling the chamber at a rate of about 0.3° C. per min. until the sample reaches about -2° C.; then (d) cooling the chamber at a rate of about 30° C. per min. until the chamber reaches a temperature of about -70° C.; then (e) warming the chamber at a rate of about 20° C. per min. until the chamber reaches a temperature of about -60° C.; then (f) holding the chamber at a temperature of about -60° C. for 17 minutes; then (g) warming the chamber at a rate of about 10° C. per min. until the chamber reaches a temperature of about -30° C.; then (h) holding the chamber at a temperature of about -30° C. for one min.; then (i) cooling the chamber at a rate of about 0.01° C. per min. until the sample reaches a temperature of about -20° C.; then (j) cooling the chamber at a rate of about 0.5° C. per min. until the sample reaches a temperature of about -65° C.; then (k) removing the blood vessel from the chamber; and then (l) placing the blood vessel in a liquid nitrogen refrigerator at a temperature of about -196° C.
References Cited (Backward Citations)
US4959319A
Cited
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1995-10-27 | 1996-04-27 | 1996-10-27 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1999-10-27 | 2000-04-27 | 2000-10-27 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 2003-10-27 | 2004-04-27 | 2004-10-27 | $7700.00 | Window Closed / Lapsed |