Patent US5723324A successfully refreshed from Google Patents.
US5723324A
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US
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Apparatus and method for electrostatic endothelial cell seeding and DNA transfection in a vascular prosthesis
Assignee: University of Akron
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Filing: 1996-09-30
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Granted: 1998-03-03
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Expires: 2016-09-30
Abstract
Abstract An apparatus and method are provided for in vitro seeding endothelial cells and transfecting DNA on the luminal wall of a vascular prosthesis. The apparatus has an internal conductor and an external conductor, and a support means which maintains a suspension containing at least one of endothelial cells and DNA plasmids in contact with the luminal wall and which maintains the luminal wall between the internal and external conductors while alteration of an electrical charge transfers at least one of endothelial cells and DNA plasmids from the suspension to the luminal wall. The internal conductor is an insulated wire and the external conductor has a bore through which the internal conductor passes. A vascular prosthesis containing a suspension of endothelial cells and/or DNA plasmids sealed within its lumen fits within the bore of the external conductor and the internal conductor wire passes through the lumen. While the prosthesis is rotated, an electrical potential is applied between the internal and external conductors altering the surface charge of the luminal wall of the prosthesis to facilitate adhesion of the endothelial cells and/or DNA plasmids to the lumen wall. When both endothelial cells and DNA plasmids are in the suspension, the altering of electric charge causes DNA plasmids within the suspension to transfect the cells. A voltage of up to 20 volts causes endothelial cells to adhere to the luminal wall and a voltage of up to 2000 volts causes DNA plasmids to transfect the endothelial cells.
Inventors
- 👤 Gary L. Bowlin
- 👤 Stanley E. Rittgers
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-07 16:36
CPC Classifications
C; C12; C12N; C12N5/00; C12N5/06; C12N5/0602; C12N5/069; A; A61; A61F; A61F2/00; A61F2/02; A61F2/04; A61F2/06; A61F2/062; A61L; A61L27/00; A61L27/50; A61L27/507; C12M; C12M25/00; C12M25/06; C12M25/08; C12M35/00; C12M35/02; Y; Y10; Y10S; Y10S623/00; Y10S623/92
Claims Analysis
Total Claims: 22
Claim 1
Independent Claim
112 words
An apparatus for modifying in vitro an entire exposed luminal wall of a synthetic vascular graft, comprising: conductor means for temporarily altering an electrical charge on the luminal wall to be modified by providing an internal conductor without openings spaced apart from an external conductor; and support means for maintaining and exposing a suspension containing at least one of endothelial cells and DNA plasmids adjacent to the entire luminal walls wherein said support means maintains the luminal wall between said internal conductor and said external conductor and wherein the alteration of the electrical charge transfers said at least one endothelial cells and DNA plasmids from the suspension to the exposed luminal wall.
Claim 2
Independent Claim
112 words
An apparatus for modifying in vitro an entire exposed luminal wall of a synthetic vascular graft, comprising: conductor means for temporarily altering an electrical charge on the luminal wall to be modified by providing an internal conductor without openings spaced apart from an external conductor; and support means for maintaining and exposing a suspension containing at least one of endothelial cells and DNA plasmids adjacent to the entire luminal walls wherein said support means maintains the luminal wall between said internal conductor and said external conductor and wherein the alteration of the electrical charge transfers said at least one endothelial cells and DNA plasmids from the suspension to the exposed luminal wall.
Claim 3
Dependent Claim
40 words
The apparatus according to claim 1, wherein said conductor means alters the electrical charge provided through the exposed luminal wall which causes endothelial cells within the suspension to adhere to the entire luminal wall of the exposed synthetic vascular graft.
Claim 4
Dependent Claim
33 words
The apparatus according to claim 2, wherein said conductor means further comprises a first voltage source for applying an electrical potential of up to about 20 volts across said internal and external conductors.
Claim 5
Dependent Claim
45 words
The apparatus according to claim 1, wherein said suspension contains both endothelial cells and DNA plasmids and wherein said conductor means alters the electric charge provided through the exposed luminal wall which causes DNA plasmids within the suspension to transfect at least the endothelial cells.
Claim 6
Dependent Claim
32 words
The apparatus according to claim 4, wherein said conductor means further comprises a voltage source for applying an electrical potential of up to about 2000 volts across said internal and external conductors.
Claim 7
Dependent Claim
82 words
The apparatus according to claim 1, wherein said conductor means comprises a pair of voltage sources for applying one of two electrical potentials across said internal and external conductors, one of said electrical potentials ranging from up to about 20 volts to cause endothelial cells in the suspension to adhere to the luminal wall and the other of said electrical potentials ranging from up to about 2000 volts to cause DNA plasmids in the suspension to transfect at least the endothelial cells.
Claim 8
Dependent Claim
43 words
The apparatus according to claim 6, wherein said external conductor is a length of electrically conductive material having a bore running the entire length thereof to support the vascular graft with the suspension received therein, said internal conductor disposed within the vascular graft.
Claim 9
Dependent Claim
15 words
The apparatus according to claim 7 further comprising drive means for rotating said external conductor.
Claim 10
Independent Claim
197 words
An apparatus for in vitro seeding endothelial cells and transfecting DNA on the luminal wall of a vascular prosthesis, the apparatus comprising: an external conductor having a bore extending the length thereof; an internal conductor adapted to be received within the vascular prosthesis which is centrally received within said bore and through which no fluid flow may occur; and a voltage source for applying an electrical potential across said external and internal conductors; whereby the vascular prosthesis is filled with a suspension comprising endothelial cells and DNA plasmid and is sealingly mounted on said internal conductor and maintained in a spaced relationship from said internal conductor within said bore of said external conductor and wherein said voltage source is adapted to apply an electrical potential across said external and internal conductors and through all of the luminal wall, wherein the luminal wall functions as a dielectric material and is altered to a more positive state, thereby temporarily altering the surface charge of the prosthesis to facilitate adhesion of the endothelial cells and transfection of the DNA plasmid thereto and wherein removal of said electrical potential causes the vascular prosthesis to return to a more negative, non-thrombogenic, state.
Claim 11
Independent Claim
197 words
An apparatus for in vitro seeding endothelial cells and transfecting DNA on the luminal wall of a vascular prosthesis, the apparatus comprising: an external conductor having a bore extending the length thereof; an internal conductor adapted to be received within the vascular prosthesis which is centrally received within said bore and through which no fluid flow may occur; and a voltage source for applying an electrical potential across said external and internal conductors; whereby the vascular prosthesis is filled with a suspension comprising endothelial cells and DNA plasmid and is sealingly mounted on said internal conductor and maintained in a spaced relationship from said internal conductor within said bore of said external conductor and wherein said voltage source is adapted to apply an electrical potential across said external and internal conductors and through all of the luminal wall, wherein the luminal wall functions as a dielectric material and is altered to a more positive state, thereby temporarily altering the surface charge of the prosthesis to facilitate adhesion of the endothelial cells and transfection of the DNA plasmid thereto and wherein removal of said electrical potential causes the vascular prosthesis to return to a more negative, non-thrombogenic, state.
Claim 12
Dependent Claim
68 words
The apparatus according to claim 9, further comprising a filling assembly for supporting said internal conductor and the prosthesis to be seeded while the prosthesis is filled and sealed with the suspension, the filling assembly having a base portion, and a pair of support blocks extending from said base portion, wherein said internal conductor may be suspended between said support blocks while the prosthesis is filled and sealed.
Claim 13
Dependent Claim
40 words
The apparatus according to claim 9, wherein said internal conductor has at least one elastomeric plug member molded thereon, said at least one elastomeric plug member having a diameter corresponding to the internal diameter of the prosthesis to be seeded.
Claim 14
Dependent Claim
80 words
The apparatus according to claim 9 further comprising: a support assembly having a base portion from which extends at least one support block, at least one bearing mount mounted on said base portion and at least one bearing adapted to rotatably journal said external conductor within said at least one bearing mount; and a drive assembly for selectively rotating the prosthesis, wherein said drive assembly includes a motor and a drive belt connected between said motor and said external conductor.
Claim 15
Dependent Claim
75 words
The apparatus according to claim 12, wherein said voltage source applies one of two electrical potentials across said internal and external conductors, one of said electrical potentials ranging from up to about 20 volts to cause endothelial cells in the solution to adhere to the luminal wall and the other of said electrical potentials ranging from up to about 2000 volts to cause DNA plasmids in the solution to transfect at least the endothelial cells.
Claim 16
Independent Claim
159 words
A method for in vitro seeding endothelial cells and transfecting DNA on the entire luminal wall of a vascular prosthesis, comprising the steps of: passing an internal conductor insulated wire without openings through the lumen of the vascular prosthesis; sealing one end of the prosthesis around the internal conductor wire; suspending the internal conductor wire and prosthesis in an upright position in a filling apparatus; filling the prosthesis with a suspension of endothelial cells and DNA plasmid; sealing the other end of the prosthesis; providing an external conductor with a bore; placing the internal conductor and prosthesis into the bore; rotating the external conductor and the prosthesis; and applying temporarily an electrical potential across the internal and external conductors and through the luminal wall while the prosthesis is being rotated such that the surface charge on the entire luminal wall is temporarily altered to facilitate adhesion of the endothelial cells and transfection of the DNA to the luminal wall.
Claim 17
Independent Claim
159 words
A method for in vitro seeding endothelial cells and transfecting DNA on the entire luminal wall of a vascular prosthesis, comprising the steps of: passing an internal conductor insulated wire without openings through the lumen of the vascular prosthesis; sealing one end of the prosthesis around the internal conductor wire; suspending the internal conductor wire and prosthesis in an upright position in a filling apparatus; filling the prosthesis with a suspension of endothelial cells and DNA plasmid; sealing the other end of the prosthesis; providing an external conductor with a bore; placing the internal conductor and prosthesis into the bore; rotating the external conductor and the prosthesis; and applying temporarily an electrical potential across the internal and external conductors and through the luminal wall while the prosthesis is being rotated such that the surface charge on the entire luminal wall is temporarily altered to facilitate adhesion of the endothelial cells and transfection of the DNA to the luminal wall.
Claim 18
Dependent Claim
81 words
The method according to claim 14, comprising the further steps of molding at least one plug member on the internal conductor wire prior to the passing of the internal conductor wire through the lumen of the prosthesis, the diameter of the plug member corresponding to the inside diameter of the vascular prosthesis; and suspending the internal conductor wire and prosthesis between a pair of support blocks while the internal conductor wire and prosthesis are inside the bore of the external conductor.
Claim 19
Dependent Claim
45 words
The method according to claim 15, wherein applying an electrical potential further comprises turning the electrical potential on for at least a period of up to 8 μsec at a voltage of up to 2000 volts to transfect the DNA plasmid into the endothelial cells.
Claim 20
Dependent Claim
47 words
The method according to claim 15, wherein applying an electrical potential further comprises turning the electrical potential on for a period no longer than 2 hours at a voltage of up to 20 volts to seed the endothelial cells to the lumen wall of the vascular prosthesis.
Claim 21
Dependent Claim
37 words
The method according to claim 15, further comprising the step of providing said suspension with a modified Dulbecco's Phosphate Buffered Saline having about 10 6 -10 7 endothelial cells/ml and about 1-2 μg/μL of linear DNA plasmids.
Claim 22
Dependent Claim
16 words
The method according to claim 18, further comprising the step of providing said suspension with antioxidants.
References Cited (Backward Citations)
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 2001-03-02 | 2001-09-01 | 2002-03-03 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 2005-03-02 | 2005-09-01 | 2006-03-03 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 2009-03-02 | 2009-09-01 | 2010-03-03 | $7700.00 | Window Closed / Lapsed |