EP0243818A2
Expired / Lapsed
EP
⏱️ 0.0 years remaining
Bioadhesives for cell and tissue adhesion
Assignee: Discovery Labware Inc
•
Filing: 1987-04-16
•
Granted: —
•
Expires: 2007-04-16
Abstract
Abstract A cell culturing system, methods for the preparation thereof, and methods for affixing other biologically active moieties to a substrate are provided. Said cell culturing system comprises: a substrate; a coating thereon of a sterile formulation comprising polyphenolic protein containing from about 35 to 100% by weight pure bioadhesive polyphenolic protein having the repeating decapeptide unit: wherein N is a whole number ranging from about 10 to about 100, wherein each X is independently selected from the group consisting of hydroxyl and hydrogen, and wherein each R is independently selected from the group consisting of hydrogen and methyl; viable cells affixed to said coated substrate; and a nutritive medium contacting said cells, whereby said cells perform normal metabolic cell functions.
Inventors
- 👤 Christine V. Benedict
- 👤 Paul T. Picciano
Bibliographic Details
Application Number
—
Jurisdiction
EP
Data Source
google_patents
Last Synchronized
2026-10-23 05:47
CPC Classifications
C; C12; C12N; C12N5/00; C12N5/0068; A; A61; A61L; A61L27/00; A61L27/36; A61L27/38; C12N2533/00; C12N2533/50
Claims Analysis
Total Claims: 70
Claim 1
Independent Claim
130 words
Method of affixing biologically active moieties to a substrate comprising (1) coating a substrate with a sterile formulation comprising polyphenolic protein containing from about 35 to 100% by weight pure bioadhesive polyphenolic protein having the repeating decapeptide unit: wherein N is a whole number ranging from about 10 to about 100, wherein each X is independently selected from the group consisting of hydroxyl and hydrogen, and wherein each R is independently selected from the group consisting of hydrogen and methyl; (2) drying said coating on said substrate; (3) fixing said coating on said substrate; (4) Rinsing said coated substrate to remove extraneous materials not firmly attached to said substrate; and (5) applying biologically active moieties to said coated substrate, whereby said biologically active moieties become affixed to said coated substrate.
Claim 2
Dependent Claim
130 words
Method of affixing biologically active moieties to a substrate comprising (1) coating a substrate with a sterile formulation comprising polyphenolic protein containing from about 35 to 100% by weight pure bioadhesive polyphenolic protein having the repeating decapeptide unit: wherein N is a whole number ranging from about 10 to about 100, wherein each X is independently selected from the group consisting of hydroxyl and hydrogen, and wherein each R is independently selected from the group consisting of hydrogen and methyl; (2) drying said coating on said substrate; (3) fixing said coating on said substrate; (4) Rinsing said coated substrate to remove extraneous materials not firmly attached to said substrate; and (5) applying biologically active moieties to said coated substrate, whereby said biologically active moieties become affixed to said coated substrate.
Claim 3
Dependent Claim
37 words
Method as defined in Claim 1 wherein said substrate is coated with from 0.1 to 2 ul of said formulation containing from about 10 to 60 µg/µl of said bioadhesive polyphenolic protein per cm 2 of substrate.
Claim 4
Dependent Claim
25 words
Method as defined in Claim 1 wherein said coating is fixed on said substrate by rinsing said substrate with an anhydrous biologically compatible sterilizing medium.
Claim 5
Dependent Claim
15 words
Method as defined in Claim 3 wherein said sterilizing medium is 30 to 100% ethanol.
Claim 6
Dependent Claim
15 words
Method as defined in Claim 3 wherein said sterilizing medium is 30 to 100% isopropanol.
Claim 7
Dependent Claim
22 words
Method as defined in Claim 1 wherein the coated substrate is rinsed to remove extraneous materials with water or a buffer solution.
Claim 8
Dependent Claim
24 words
Method as defined in Claim 1 wherein said substrate is selected from the group consisting of synthetic polymeric materials, glass, metals, and microporous filters.
Claim 9
Dependent Claim
11 words
Method as defined in Claim 7 wherein said substrate is polytetrafluoroethylene.
Claim 10
Dependent Claim
139 words
Method of affixing viable cells to a substrate comprising (1) coating a substrate with a sterile formulation comprising polyphenolic protein containing from about 35 to 100% by weight pure bioadhesive polyphenolic protein having the repeating decapeptide unit: wherein N is a whole number ranging from about 10 to about 100, wherein each X is independently selected from the group consisting of hydroxyl and hydrogen, and wherein each R is independently selected from the group consisting of hydrogen and methyl; (2) drying said coating on said substrate; (3) fixing said coating on said substrate; (4) rinsing said coated substrate to remove extraneous materials not firmly attached to said substrate; and (5) applying viable cells to said coated substrate, whereby said cells become affixed to said coated substrate and, in the presence of a nutritive environment, perform normal metabolic cell functions.
Claim 11
Dependent Claim
139 words
Method of affixing viable cells to a substrate comprising (1) coating a substrate with a sterile formulation comprising polyphenolic protein containing from about 35 to 100% by weight pure bioadhesive polyphenolic protein having the repeating decapeptide unit: wherein N is a whole number ranging from about 10 to about 100, wherein each X is independently selected from the group consisting of hydroxyl and hydrogen, and wherein each R is independently selected from the group consisting of hydrogen and methyl; (2) drying said coating on said substrate; (3) fixing said coating on said substrate; (4) rinsing said coated substrate to remove extraneous materials not firmly attached to said substrate; and (5) applying viable cells to said coated substrate, whereby said cells become affixed to said coated substrate and, in the presence of a nutritive environment, perform normal metabolic cell functions.
Claim 12
Dependent Claim
38 words
Method as defined in Claim 9 wherein said substrate is coated with from 0.1 to 2 µl of said formulation containing from about 10 to 60 µg/ µl of said bioadhesive polyphenolic protein per cm 2 of substrate.
Claim 13
Dependent Claim
25 words
Method as defined in Claim 9 wherein said coating is fixeo on said substrate by rinsing said substrate with an anhydrous biologically compatible sterilizing medium.
Claim 14
Dependent Claim
16 words
Method as defined in Claim 11 wherein said sterilizing medium is 30 to 100 % ethanol.
Claim 15
Dependent Claim
15 words
13 . Method as defined in Claim 11 wherein said sterilizing medium is 30-100% isopropanol.
Claim 16
Dependent Claim
23 words
Method as defined in Claim 9 wherein the coated substrate is rinsed to remove extraneous materials with sterile aqueous serum-free tissue culture medium.
Claim 17
Dependent Claim
21 words
Method as defined in Claim 9 wherein the coated substrate is rinsed to remove extraneous materials with water or buffer solution.
Claim 18
Dependent Claim
20 words
Method as defined in Claim 9 wherein the viable cells are applied to the coated substrate in a serum-containing medium.
Claim 19
Dependent Claim
20 words
Method as defined in Claim 9 wherein the viable cells are applied to said coated substrate in a serum-free medium.
Claim 20
Dependent Claim
14 words
Method as defined in Claim 9 wherein said substrate is an in vitro substrate.
Claim 21
Dependent Claim
24 words
Method as defined in Claim 18 wherein said substrate is selected from the group consisting of synthetic polymeric materials, glass, metals, and microporous filters.
Claim 22
Dependent Claim
11 words
Method as defined in Claim 19 wherein said substrate is polytetrafluoroethylene.
Claim 23
Dependent Claim
14 words
Method as defined in Claim 9 wherein said substrate is an in vivo substrate.
Claim 24
Dependent Claim
24 words
Method as defined in Claim 21 wherein the substrate is selected from the group consisting of collagen, laminin, fibronectin, prosthetic grafting materials and polylysine.
Claim 25
Dependent Claim
11 words
Method as defined in Claim 22 wherein said substrate is polytetrafluoroethylene.
Claim 26
Dependent Claim
98 words
A cell culturing system comprising: a substrate; a coating thereon of a sterile formultion comprising polyphenolic protein containing from about 35 to 100% by weight pure bioadhesive polyphenolic protein having the repeating decapeptide unit: wherein N is a whole number ranging from about 10 to about 100, wherein each X is independently selected from the group consisting of hydroxyl and hydrogen, and wherein each R is independently selected from the group consisting of hydrogen and methyl; viable cells affixed' to said coated substrate; and a nutritive medium contacting said cells, whereby said cells perform normal metabolic cell functions.
Claim 27
Dependent Claim
98 words
A cell culturing system comprising: a substrate; a coating thereon of a sterile formultion comprising polyphenolic protein containing from about 35 to 100% by weight pure bioadhesive polyphenolic protein having the repeating decapeptide unit: wherein N is a whole number ranging from about 10 to about 100, wherein each X is independently selected from the group consisting of hydroxyl and hydrogen, and wherein each R is independently selected from the group consisting of hydrogen and methyl; viable cells affixed' to said coated substrate; and a nutritive medium contacting said cells, whereby said cells perform normal metabolic cell functions.
Claim 28
Dependent Claim
17 words
A cell culturing system as defined in Claim 24 wherein the substrate is an in vitro substrate.
Claim 29
Dependent Claim
27 words
A cell culturing system as defined in Claim 25 wherein the substrate is selected from the group consisting of synthetic polymeric material, glass, metals, and microporous filters.
Claim 30
Dependent Claim
14 words
A cell culturing system as defined in Claim 26 wherein said substrate is polytetrafluoroethylene.
Claim 31
Dependent Claim
18 words
. A cell culturing system as defined in Claim 24 wherein said substrate is an in vivo substrate.
Claim 32
Dependent Claim
27 words
A cell culturing system as defined in Claim 28 wherein the substrate is selected from the group consisting of collagen, laminin, fibronectin, prosthetic grafting materials and polylysine.
Claim 33
Dependent Claim
14 words
A cell culturing system as defined in Claim 29 wherein the substrate is polytetrafluoroethylene.
Claim 34
Dependent Claim
123 words
Method of affixing biologically active moieties to a substrate, comprising: (1) dispersing said biologically active moieties in a serum-free solution; (2) admixing a sterile formulation comprising polyphenolic protein containing from about 35 to 100% by weight pure bioadhesive polyphenolic protein having the repeating decapeptide unit: wherein N is a whole number ranging from about 10 to about 100, wherein each X is independently selected from the group consisting of hydroxyl and hydrogen, and wherein each R is independently selected from the group consisting of hydrogen and methyl; with said dispersion of biologically active moieties, thereby attaching said bioadhesive polyphenolic protein to said biologically active moieties; (3) recovering the resulting biologically active moieties; and (4) affixing said recovered biologically active moieties to said substrate.
Claim 35
Dependent Claim
123 words
Method of affixing biologically active moieties to a substrate, comprising: (1) dispersing said biologically active moieties in a serum-free solution; (2) admixing a sterile formulation comprising polyphenolic protein containing from about 35 to 100% by weight pure bioadhesive polyphenolic protein having the repeating decapeptide unit: wherein N is a whole number ranging from about 10 to about 100, wherein each X is independently selected from the group consisting of hydroxyl and hydrogen, and wherein each R is independently selected from the group consisting of hydrogen and methyl; with said dispersion of biologically active moieties, thereby attaching said bioadhesive polyphenolic protein to said biologically active moieties; (3) recovering the resulting biologically active moieties; and (4) affixing said recovered biologically active moieties to said substrate.
Claim 36
Dependent Claim
130 words
Method of affixing biologically active moieties to a substrate comprising (1) coating a substrate with a sterile formulation comprising polyphenolic protein containing from about 35 to 100% by weight pure bioadhesive polyphenolic protein having the repeating decapeptide unit: wherein N is a whole number ranging from about 10 to about 100, wherein each X is independently selected from the group consisting of hydroxyl and hydrogen, and wherein each R is independently selected from the group consisting of hydrogen and methyl; (2) drying said coating on said substrate; (3) fixing said coating on said substrate; (4) Rinsing said coated substrate to remove extraneous materials not firmly attached to said substrate; and (5) applying biologically active moieties to said coated substrate, whereby said biologically active moieties become affixed to said coated substrate.
Claim 37
Dependent Claim
130 words
Method of affixing biologically active moieties to a substrate comprising (1) coating a substrate with a sterile formulation comprising polyphenolic protein containing from about 35 to 100% by weight pure bioadhesive polyphenolic protein having the repeating decapeptide unit: wherein N is a whole number ranging from about 10 to about 100, wherein each X is independently selected from the group consisting of hydroxyl and hydrogen, and wherein each R is independently selected from the group consisting of hydrogen and methyl; (2) drying said coating on said substrate; (3) fixing said coating on said substrate; (4) Rinsing said coated substrate to remove extraneous materials not firmly attached to said substrate; and (5) applying biologically active moieties to said coated substrate, whereby said biologically active moieties become affixed to said coated substrate.
Claim 38
Dependent Claim
37 words
Method as defined in Claim 1 wherein said substrate is coated with from 0.1 to 2 ul of said formulation containing from about 10 to 60 µg/µl of said bioadhesive polyphenolic protein per cm 2 of substrate.
Claim 39
Dependent Claim
25 words
Method as defined in Claim 1 wherein said coating is fixed on said substrate by rinsing said substrate with an anhydrous biologically compatible sterilizing medium.
Claim 40
Dependent Claim
15 words
Method as defined in Claim 3 wherein said sterilizing medium is 30 to 100% ethanol.
Claim 41
Dependent Claim
15 words
Method as defined in Claim 3 wherein said sterilizing medium is 30 to 100% isopropanol.
Claim 42
Dependent Claim
22 words
Method as defined in Claim 1 wherein the coated substrate is rinsed to remove extraneous materials with water or a buffer solution.
Claim 43
Dependent Claim
24 words
Method as defined in Claim 1 wherein said substrate is selected from the group consisting of synthetic polymeric materials, glass, metals, and microporous filters.
Claim 44
Dependent Claim
11 words
Method as defined in Claim 7 wherein said substrate is polytetrafluoroethylene.
Claim 45
Dependent Claim
139 words
Method of affixing viable cells to a substrate comprising (1) coating a substrate with a sterile formulation comprising polyphenolic protein containing from about 35 to 100% by weight pure bioadhesive polyphenolic protein having the repeating decapeptide unit: wherein N is a whole number ranging from about 10 to about 100, wherein each X is independently selected from the group consisting of hydroxyl and hydrogen, and wherein each R is independently selected from the group consisting of hydrogen and methyl; (2) drying said coating on said substrate; (3) fixing said coating on said substrate; (4) rinsing said coated substrate to remove extraneous materials not firmly attached to said substrate; and (5) applying viable cells to said coated substrate, whereby said cells become affixed to said coated substrate and, in the presence of a nutritive environment, perform normal metabolic cell functions.
Claim 46
Dependent Claim
139 words
Method of affixing viable cells to a substrate comprising (1) coating a substrate with a sterile formulation comprising polyphenolic protein containing from about 35 to 100% by weight pure bioadhesive polyphenolic protein having the repeating decapeptide unit: wherein N is a whole number ranging from about 10 to about 100, wherein each X is independently selected from the group consisting of hydroxyl and hydrogen, and wherein each R is independently selected from the group consisting of hydrogen and methyl; (2) drying said coating on said substrate; (3) fixing said coating on said substrate; (4) rinsing said coated substrate to remove extraneous materials not firmly attached to said substrate; and (5) applying viable cells to said coated substrate, whereby said cells become affixed to said coated substrate and, in the presence of a nutritive environment, perform normal metabolic cell functions.
Claim 47
Dependent Claim
37 words
Method as defined in Claim 9 wherein said substrate is coated with from 0.1 to 2pl of said formulation containing from about 10 to 60 µg/ µl of said bioadhesive polyphenolic protein per cm 2 of substrate.
Claim 48
Dependent Claim
25 words
Method as defined in Claim 9 wherein said coating is fixed on said substrate by rinsing said substrate with an anhydrous biologically compatible sterilizing medium.
Claim 49
Dependent Claim
15 words
Method as defined in Claim 11 wherein said sterilizing medium is 30 to 100% ethanol.
Claim 50
Dependent Claim
13 words
Method as defined in Claim 11 wherein said sterilizing medium is 30-100% isopropanol.
Claim 51
Dependent Claim
23 words
Method as defined in Claim 9 wherein the coated substrate is rinsed to remove extraneous materials with sterile aqueous serum-free tissue culture medium.
Claim 52
Dependent Claim
21 words
Method as defined in Claim 9 wherein the coated substrate is rinsed to remove extraneous materials with water or buffer solution.
Claim 53
Dependent Claim
20 words
Method as defined in Claim 9 wherein the viable cells are applied to the coated substrate in a serum-containing medium.
Claim 54
Dependent Claim
20 words
Method as defined in Claim 9 wherein the viable cells are applied to said coated substrate in a serum-free medium.
Claim 55
Dependent Claim
14 words
Method as defined in Claim 9 wherein said substrate is an in vitro substrate.
Claim 56
Dependent Claim
24 words
Method as defined in Claim 18 wherein said substrate is selected from the group consisting of synthetic polymeric materials, glass, metals, and microporous filters.
Claim 57
Dependent Claim
11 words
Method as defined in Claim 19 wherein said substrate is polytetrafluoroethylene.
Claim 58
Dependent Claim
14 words
Method as defined in Claim 9 wherein said substrate is an in vivo substrate.
Claim 59
Dependent Claim
24 words
Method as defined in Claim 21 wherein the substrate is selected from the group consisting of collagen, laminin, fibronectin, prosthetic grafting materials and polylysine.
Claim 60
Dependent Claim
11 words
Method as defined in Claim 22 wherein said substrate is polytetrafluoroethylene.
Claim 61
Dependent Claim
102 words
Use of a system comprising: a substrate; a coating thereon of a sterile formultion comprising polyphenolic protein containing from about 35 to 100 % by weight pure bioadhesive polyphenolic protein having the repeating decapeptide unit: wherein N is a whole number ranging from about 10 to about 100, wherein each X is independently selected from the group consisting of hydroxyl and hydrogen, and wherein each R is independently selected from the group consisting of hydrogen and methyl; viable cells affixed to said coated substrate; and a nutritive medium contacting said cells, for culturing cells wherein said cells perform normal metabolic cell functions.
Claim 62
Dependent Claim
102 words
Use of a system comprising: a substrate; a coating thereon of a sterile formultion comprising polyphenolic protein containing from about 35 to 100 % by weight pure bioadhesive polyphenolic protein having the repeating decapeptide unit: wherein N is a whole number ranging from about 10 to about 100, wherein each X is independently selected from the group consisting of hydroxyl and hydrogen, and wherein each R is independently selected from the group consisting of hydrogen and methyl; viable cells affixed to said coated substrate; and a nutritive medium contacting said cells, for culturing cells wherein said cells perform normal metabolic cell functions.
Claim 63
Dependent Claim
19 words
Use of the system as defined in claim 24, wherein an in vitro substrate is used as said substrate.
Claim 64
Dependent Claim
28 words
Use of the system as defined in claim 25, wherein a substrate selected from the group consisting of synthetic polymeric material, glass, metals, and microporous filters is used.
Claim 65
Dependent Claim
16 words
Use of the system as defined in claim 26, wherein a substrate of polytetrafluoroethylene is used.
Claim 66
Dependent Claim
19 words
Use of the system as defined in claim 24, wherein an in vivo substrate is used as said substrate.
Claim 67
Dependent Claim
28 words
Use of the system as defined in claim 28, wherein a substrate selected from the group consisting of collagen, laminin, fibronectin, prosthetic grafting materials and polylysine is used.
Claim 68
Dependent Claim
16 words
Use of the system as defined in claim 29, wherein a substrate of polytetrafluoroethylene is used.
Claim 69
Dependent Claim
122 words
Method of affixing biologically active moieties to a substrate, comprising: (1) dispersing said biologically active moieties in a serum-free solution; (2) admixing a sterile formulation comprising polyphenolic protein containing from about 35 to 100% by-weight pure bioadhesive polyphenolic protein having the repeating decapeptide unit: wherein N is a whole number ranging from about 10 to about 100, wherein each X is independently selected from the group consisting of hydroxyl and hydrogen, and wherein each R is independently selected from the group consisting of hydrogen and methyl; with said dispersion of biologically active moieties, thereby attaching said bioadhesive polyphenolic protein to said biologically active moieties; (3) recovering the resulting biologically active moieties; and (4) affixing said recovered biologically active moieties to said substrate.
Claim 70
Dependent Claim
122 words
Method of affixing biologically active moieties to a substrate, comprising: (1) dispersing said biologically active moieties in a serum-free solution; (2) admixing a sterile formulation comprising polyphenolic protein containing from about 35 to 100% by-weight pure bioadhesive polyphenolic protein having the repeating decapeptide unit: wherein N is a whole number ranging from about 10 to about 100, wherein each X is independently selected from the group consisting of hydroxyl and hydrogen, and wherein each R is independently selected from the group consisting of hydrogen and methyl; with said dispersion of biologically active moieties, thereby attaching said bioadhesive polyphenolic protein to said biologically active moieties; (3) recovering the resulting biologically active moieties; and (4) affixing said recovered biologically active moieties to said substrate.
References Cited (Backward Citations)
No citation records indexed.
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