Patent US6080579A successfully refreshed from Google Patents.
US6080579A
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Method for producing human intervertebral disc cells
Assignee: Carolinas Healthcare System
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Filing: 1997-11-26
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Granted: 2000-06-27
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Expires: 2017-11-26
Abstract
Abstract There is provided a method for growing human intervertebral cells. Disc tissue is surgically removed from a normal disc of a patient, the cells expanded by feeding with a cell stimulant such as a growth factor, or a cytokine or a bioactive agent to form monolayer primary cell cultures on a plastic mesh such as a nylon mesh. In the case of a growth factor, fetal bovine serum is preferred as it improves cell proliferation and production of appropriate extracellular matrix components. In another aspect of this invention, the monolayer primary cell cultures are seeded in alginate or agarose and fed again with the cell stimulant until three-dimensional cell cultures are formed. The cells are recovered from the alginate or agarose or from monolayer cultures. Re-implantation is carried out using bioresorbable carriers or cell suspensions.
Inventors
- 👤 Edward Nathaniel Hanley, Jr.
- 👤 Helen Elizabeth Gruber
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-23 19:34
CPC Classifications
C; C12; C12N; C12N5/00; C12N5/06; C12N5/0602; C12N5/0652; C12N5/0655; A; A61; A61P; A61P19/00; A61P43/00; C12N5/0068; A61K; A61K35/00; A61K35/12; A61K38/00; C12N2500/00; C12N2500/30; C12N2500/32; C12N2501/00; C12N2501/10; C12N2501/15; C12N2533/00; C12N2533/30; C12N2533/70; C12N2533/74
Claims Analysis
Total Claims: 13
Claim 1
Independent Claim
70 words
A method for propagating human intervertebral disc cells in vitro comprising: a) providing human intervertebral disc tissue; b) mincing said human intervertebral disc tissue to obtain a minced explant comprising human intervertebral disc cells; and c) culturing said minced explant under conditions to propagate and form a monolayer of human intervertebral disc cells, wherein the human intervertebral disc cells of said monolayer can be isolated and further propagated upon passaging.
Claim 2
Independent Claim
70 words
A method for propagating human intervertebral disc cells in vitro comprising: a) providing human intervertebral disc tissue; b) mincing said human intervertebral disc tissue to obtain a minced explant comprising human intervertebral disc cells; and c) culturing said minced explant under conditions to propagate and form a monolayer of human intervertebral disc cells, wherein the human intervertebral disc cells of said monolayer can be isolated and further propagated upon passaging.
Claim 3
Dependent Claim
15 words
The method of claim 1, wherein said minced explant is anchored by a plastic mesh.
Claim 4
Dependent Claim
50 words
A human intervertebral disc cell culture obtained by the method of claim 2 comprising a monolayer of human intervertebral disc cells and a human intervertebral disc minced explant, wherein at least a portion of said monolayer of human intervertebral disc cells is attached to said human intervertebral disc minced explant.
Claim 5
Dependent Claim
20 words
The method of claim 1 wherein said minced explant is cultured in the presence of serum, growth factors or cytokines.
Claim 6
Dependent Claim
18 words
The method of claim 4 wherein said minced explant is cultured in the presence of fetal bovine serum.
Claim 7
Dependent Claim
20 words
The method of claim 4 wherein said minced explant is cultured in the presence of transforming growth factor beta (TGF-β).
Claim 8
Dependent Claim
50 words
A human intervertebral disc cell culture obtained by the method of claim 1 comprising a monolayer of human intervertebral disc cells and a human intervertebral disc minced explant, wherein at least a portion of said monolayer of human intervertebral disc cells is attached to said human intervertebral disc minced explant.
Claim 9
Independent Claim
120 words
A method for culturing human intervertebral disc cells in a three-dimensional structure in vitro comprising: a) providing human intervertebral disc tissue; b) mincing said human intervertebral disc tissue to obtain a minced explant comprising human intervertebral disc cells; c) culturing said minced explant under conditions to propagate and form a monolayer of human intervertebral disc cells, wherein the human intervertebral disc cells of said monolayer can be isolated and further propagated upon passaging; d) isolating said human intervertebral disc cells from said monolayer; e) seeding the isolated cells into a carrier such that the isolated cells are dispersed and distributed in the carrier thereby forming a three-dimensional structure; and f) culturing said dispersed and distributed cells in the three-dimensional structure.
Claim 10
Independent Claim
120 words
A method for culturing human intervertebral disc cells in a three-dimensional structure in vitro comprising: a) providing human intervertebral disc tissue; b) mincing said human intervertebral disc tissue to obtain a minced explant comprising human intervertebral disc cells; c) culturing said minced explant under conditions to propagate and form a monolayer of human intervertebral disc cells, wherein the human intervertebral disc cells of said monolayer can be isolated and further propagated upon passaging; d) isolating said human intervertebral disc cells from said monolayer; e) seeding the isolated cells into a carrier such that the isolated cells are dispersed and distributed in the carrier thereby forming a three-dimensional structure; and f) culturing said dispersed and distributed cells in the three-dimensional structure.
Claim 11
Dependent Claim
16 words
The method of claim 8, wherein said three-dimensional structure is formed in a cell well insert.
Claim 12
Dependent Claim
10 words
The method of claim 8, wherein said carrier is alginate.
Claim 13
Dependent Claim
10 words
The method of claim 8, wherein said carrier is agarose.
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USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 2003-06-27 | 2003-12-27 | 2004-06-27 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 2007-06-27 | 2007-12-27 | 2008-06-27 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 2011-06-27 | 2011-12-27 | 2012-06-27 | $7700.00 | Window Closed / Lapsed |