US4820626A
Expired / Lapsed
US
⏱️ 0.0 years remaining
Method of treating a synthetic or naturally occuring surface with microvascular endothelial cells, and the treated surface itself
Assignee: Thomas Jefferson University
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Filing: 1985-06-06
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Granted: 1989-04-11
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Expires: 2005-06-06
Abstract
Abstract The present invention provides a novel method of treating a synthetic or naturally occurring implant, such as a vascular graft, intended for implantation in a human patient, comprising obtaining human microvascular rich tissue from that patient; separating microvascular endothelial cells from that tissue; and placing said microvascular endothelial cells onto said implant to provide at least about 50% confluence of said cells on the surface of said implant to be treated. In the preferred embodiment, the microvascular rich tissue is perinephric fat, which is obtained from the donor and subjected to a digestive separation process to provide an abundant supply of microvascular endothelial cells. In the preferred embodiment, these microvascular endothelial cells are placed onto the implant by suspending them in a protein (plasma) containing physiologic saline solution, which is incubated on the graft until the endothelial cells either adhere to the graft surface or clot within the protein solution. An efficient method is therefore provided which will permit a patient to donate excess microvascularized tissue which may be used to procure autologous endothelial cells which are subsequently used to treat a graft or other implant to provide at least 50% confluence while his related surgical procedure is in progress.
Inventors
- 👤 Stuart K. Williams
- 👤 Bruce E. Jarrell
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-23 00:09
CPC Classifications
A; A61; A61L; A61L27/00; A61L27/50; A61L27/507; A61L27/28; A61L27/34
Claims Analysis
Total Claims: 39
Claim 1
Independent Claim
68 words
A method of treating an implant intended for implantation in a human patient, comprising: obtaining human microvascular rich tissue from that patient; separating microvascular endothelial cells from said patient's tissue; applying said separated uncultured microvascular endothelial cells onto said implant to provide at least about 50% confluence of said cells on the surface of said implant to be treated; and implanting said implant into a patient without culturing.
Claim 2
Independent Claim
68 words
A method of treating an implant intended for implantation in a human patient, comprising: obtaining human microvascular rich tissue from that patient; separating microvascular endothelial cells from said patient's tissue; applying said separated uncultured microvascular endothelial cells onto said implant to provide at least about 50% confluence of said cells on the surface of said implant to be treated; and implanting said implant into a patient without culturing.
Claim 3
Dependent Claim
11 words
The method of claim 1 wherein said tissue is fat tissue.
Claim 4
Dependent Claim
12 words
The method of claim 2 wherein said fat tissue is perinephric fat.
Claim 5
Dependent Claim
12 words
The method of claim 2 wherein said fat tissue is subcutaneous fat.
Claim 6
Dependent Claim
10 words
The method of claim 1 wherein said tissue is omentum.
Claim 7
Dependent Claim
10 words
The method of claim 1 wherein said tissue is intrathoracic.
Claim 8
Dependent Claim
10 words
The method of claim 1 wherein said tissue is intraperitoneal.
Claim 9
Dependent Claim
16 words
The method of claim 1 wherein said method further comprises implanting said implant to said donor.
Claim 10
Dependent Claim
18 words
The method of claim 8 wherein said method is performed in its entirety in a single sterile environment.
Claim 11
Dependent Claim
12 words
The method of claim 1 wherein said implant is a vascular graft.
Claim 12
Dependent Claim
13 words
The method of claim 10 wherein said vascular graft is a polyester graft.
Claim 13
Dependent Claim
15 words
The method of claim 1 wherein said separating step is conducted using a proteolytic enzyme.
Claim 14
Dependent Claim
12 words
The method of claim 12 wherein said proteolytic enzyme is a collagenase.
Claim 15
Dependent Claim
13 words
The method of claim 13 wherein said proteolytic enzyme comprises caseanase and trypsin.
Claim 16
Dependent Claim
27 words
The method of claim 12 wherein said separating step comprises incubating said tissue with said proteolytic enzyme until this tissue mass disperses to produce a tissue digest.
Claim 17
Dependent Claim
14 words
The method of claim 13 wherein microvascular endothelial cells are separated from said digest.
Claim 18
Dependent Claim
23 words
The method of claim 16 wherein said microvascular endothelial cells are separated by low speed centrifugation to produce an endothelial cell rich pellet.
Claim 19
Dependent Claim
15 words
The method of claim 17 wherein said pellet is washed with a buffered saline solution.
Claim 20
Dependent Claim
34 words
The method of claim 16 wherein said endothelial cells are purified from said pellet by centrifugation in a continuous gradient and harvesting of the endothelial cell band or by selective sieving of endothelial cells.
Claim 21
Dependent Claim
20 words
The method of claim 1 wherein said applying step comprises suspending said microvascular endothelial cells in a buffered saline solution.
Claim 22
Dependent Claim
20 words
The method of claim 20 wherein said suspending step comprises suspending in a buffered saline solution comprising about 1% protein.
Claim 23
Dependent Claim
42 words
The method of claim 20 wherein said separating step comprises obtaining a microvascular endothelial cell rich pellet, and wherein said suspending step comprises suspending said pellets in said buffered saline solution in a pellet to solution volumetric ratio of 1:5 to 1:15.
Claim 24
Dependent Claim
11 words
The method of claim 22 wherein said ratio is about 1:10.
Claim 25
Dependent Claim
27 words
The method of claim 1 wherein said microvascular endothelial cells are applied in suspension in a physiologic solution, to the surface of the implant to be treated.
Claim 26
Dependent Claim
15 words
The method of claim 24 wherein said surface of said implant is pretreated with protein.
Claim 27
Dependent Claim
11 words
The method of claim 25 wherein said protein is plasma protein.
Claim 28
Dependent Claim
14 words
The method of claim 26 wherein said plasma protein is said human patient's plasma.
Claim 29
Dependent Claim
22 words
The method of claim 26 wherein said protein pretreatment comprises applying platelet rich plasma or blood to said surface of said implant.
Claim 30
Dependent Claim
24 words
The method of claim 24 wherein said microvascular endothelial cells are incubated on said surface until adherence to provide at least 50% confluence occurs.
Claim 31
Dependent Claim
16 words
The method of claim 29 wherein said application provides confluent or near confluent endothelial cell coverage.
Claim 32
Dependent Claim
20 words
The method of claim 24 wherein said microvascular endothelial cells are provided at a rate in excess of 100% confluence.
Claim 33
Dependent Claim
19 words
The method of claim 24 wherein said surface of said implant has been pretreated to emulate a protein surface.
Claim 34
Dependent Claim
20 words
The method of claim 24 wherein said microvascular endothelial cells are preclotted in a protein gel applied to said graft.
Claim 35
Dependent Claim
13 words
The method of claim 33 wherein said protein gel is a fibrin gel.
Claim 36
Dependent Claim
15 words
The method of claim 33 herein said microvascular endothelial cells are preclotted in human plasma.
Claim 37
Dependent Claim
12 words
The method of claim 35 wherein said plasma is said patient's plasma.
Claim 38
Dependent Claim
15 words
The method of claim 1 wherein said implant is selected to be a vascular graft.
Claim 39
Dependent Claim
12 words
The method of claim 37 wherein said graft is a polyester graft.
References Cited (Backward Citations)
US3276448A
Cited
WO1983003536A1
Cited
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