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US5512474A
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US
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Cell culture support containing a cell adhesion factor and a positively-charged molecule
Assignee: Surmodics IVD Inc
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Filing: 1994-03-09
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Granted: 1996-04-30
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Expires: 2014-03-09
Abstract
Abstract A combination of a cell adhesion factor and a positively-charged molecule are bound to the surface of a cell culture support of a bioreactor to improve cell attachment and stabilize cell growth. The positively charged molecule is preferably polylysine, chitosan, poly(ethyleneimine) or acrylics polymerized from acrylamide or methacrylamide and incorporating positively-charged groups in the form of primary, secondary or tertiary amines, or quaternary salts. The cell adhesion factor is preferably fibronectin, laminin, collagen, vitronectin or tenascin, or fragments or analogs having a cell binding domain thereof. The positively-charged molecule and the cell adhesion factor can be covalently bound to the supporting surface. In another embodiment, the positively-charged molecule and the cell adhesion factor are covalently bound to one another and either the positively-charged molecule or the cell adhesion factor is covalently bound to the supporting surface. Also, the positively-charged molecule or the cell adhesion factor or both cam be provided in the form of a stable coating non-covalently bound around the surface of the support. The cell culture support may be in the form of a microcarrier and can be made of polystyrene or polypropylene
Inventors
- 👤 David L. Clapper
- 👤 Wei-Shou Hu
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-23 19:21
CPC Classifications
C; C12; C12N; C12N5/00; C12N5/0068; C12N5/0075; C12M; C12M25/00; C12M25/16; C12N2533/00; C12N2533/30; C12N2533/32; C12N2533/50; C12N2533/54; C12N2533/70; Y; Y10; Y10S; Y10S530/00; Y10S530/81; Y10S530/812; Y10S530/815
Claims Analysis
Total Claims: 14
Claim 1
Independent Claim
176 words
A cell culture support comprising a support material in the form of a microcarrier and comprising a supporting surface for the attachment of cells, the surface bearing a combination comprising: a positively-charged molecule selected from a group consisting of polylysine, chitosan, poly(ethyleneimine), and acrylics polymerized from acrylamide or methacrylamide and incorporating positively-charged groups in the form of primary, secondary, or tertiary amines, or quaternary salts, and a cell adhesion factor selected from the group consisting of fibronectin, laminin, collagen, vitronectin and tenascin, and active fragments and synthetic analogs having a cell binding domain thereof, wherein either (a) the positively-charged molecule and the cell adhesion factor are covalently bound to the supporting surface, or (b) the positively-charged molecule and the cell adhesion factor are covalently bound to one another and either the positively-charged molecule or the cell adhesion factor is covalently bound to the supporting surface, or (c) either the positively-charged molecule or the cell adhesion factor, or both, are provided in the form of a stable coating noncovalently bound around the surface of the supporting material.
Claim 2
Dependent Claim
176 words
A cell culture support comprising a support material in the form of a microcarrier and comprising a supporting surface for the attachment of cells, the surface bearing a combination comprising: a positively-charged molecule selected from a group consisting of polylysine, chitosan, poly(ethyleneimine), and acrylics polymerized from acrylamide or methacrylamide and incorporating positively-charged groups in the form of primary, secondary, or tertiary amines, or quaternary salts, and a cell adhesion factor selected from the group consisting of fibronectin, laminin, collagen, vitronectin and tenascin, and active fragments and synthetic analogs having a cell binding domain thereof, wherein either (a) the positively-charged molecule and the cell adhesion factor are covalently bound to the supporting surface, or (b) the positively-charged molecule and the cell adhesion factor are covalently bound to one another and either the positively-charged molecule or the cell adhesion factor is covalently bound to the supporting surface, or (c) either the positively-charged molecule or the cell adhesion factor, or both, are provided in the form of a stable coating noncovalently bound around the surface of the supporting material.
Claim 3
Dependent Claim
14 words
A support according to claim 1 wherein the surface bears the combination of (a).
Claim 4
Dependent Claim
14 words
A support according to claim 1 wherein the surface bears the combination of (b).
Claim 5
Dependent Claim
14 words
A support according to claim 1 wherein the surface bears the combination of (c).
Claim 6
Dependent Claim
20 words
A support according to claim 1 wherein the support material is selected from the group consisting of polystyrene and polypropylene.
Claim 7
Dependent Claim
12 words
A support according to claim 1 wherein the support material is polystyrene.
Claim 8
Independent Claim
187 words
A method of preparing a cell culture support comprising the steps of: providing a support material in the form of a microcarrier and comprising a support surface for the attachment of cells and applying to the support surface a combination comprising: a positively-charged molecule selected from a group consisting of polylysine, chitosan, poly(ethyleneimine), and, acrylics polymerized from acrylamide or methacrylamide and incorporating positively-charged groups in the form of primary, secondary, or tertiary amines, or quaternary salts, and a cell adhesion factor selected from the group consisting of fibronectin, laminin, collagen, vitronectin and tenascin, and active fragments and synthetic analogs having a cell binding domain thereof, wherein either (a) the positively-charged molecule and the cell adhesion factor are covalenfly bound to the supporting surface, or (b) the positively-charged molecule and the cell adhesion factor are covalently bound to one another and either the positively-charged molecule or the cell adhesion factor is covalently bound to the supporting surface, or (c) either the positively-charged molecule or the cell adhesion factor, or both, are provided in the form of a stable coating noncovalently bound around the surface of the supporting material.
Claim 9
Independent Claim
187 words
A method of preparing a cell culture support comprising the steps of: providing a support material in the form of a microcarrier and comprising a support surface for the attachment of cells and applying to the support surface a combination comprising: a positively-charged molecule selected from a group consisting of polylysine, chitosan, poly(ethyleneimine), and, acrylics polymerized from acrylamide or methacrylamide and incorporating positively-charged groups in the form of primary, secondary, or tertiary amines, or quaternary salts, and a cell adhesion factor selected from the group consisting of fibronectin, laminin, collagen, vitronectin and tenascin, and active fragments and synthetic analogs having a cell binding domain thereof, wherein either (a) the positively-charged molecule and the cell adhesion factor are covalenfly bound to the supporting surface, or (b) the positively-charged molecule and the cell adhesion factor are covalently bound to one another and either the positively-charged molecule or the cell adhesion factor is covalently bound to the supporting surface, or (c) either the positively-charged molecule or the cell adhesion factor, or both, are provided in the form of a stable coating noncovalently bound around the surface of the supporting material.
Claim 10
Dependent Claim
14 words
A method according to claim 7 wherein the surface bears the combination of (a).
Claim 11
Dependent Claim
14 words
A support according to claim 7 wherein the surface bears the combination of (b).
Claim 12
Dependent Claim
14 words
A support according to claim 7 wherein the surface bears the combination of (c).
Claim 13
Dependent Claim
20 words
A support according to claim 7 wherein the support material is selected from the group consisting of polystyrene and polypropylene.
Claim 14
Dependent Claim
12 words
A support according to claim 7 wherein the support material is polystyrene.
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) | 1999-04-30 | 1999-10-30 | 2000-04-30 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 2003-04-30 | 2003-10-30 | 2004-04-30 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 2007-04-30 | 2007-10-30 | 2008-04-30 | $7700.00 | Window Closed / Lapsed |