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Patent US4559304A successfully refreshed from Google Patents.
US4559304A
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Substratum for cell culture and a method for culturing and isolating cells using same
Assignee: Koken Co Ltd
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Filing: 1983-08-08
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Granted: 1985-12-17
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Expires: 2003-08-08
Abstract
Abstract A substratum for cell culture which comprises a chemically modified collagen rich in either positive or negative charges when under culture conditions. The substratum is prepared by modifying the amino groups or carboxyl groups of collagen. The chemically modified collagen enhances the adherence and proliferation of animal cells much more actively than unmodified collagen in the presence or absence of bovine fetus serum. The cultured animal cells can be detached efficiently from the chemically modified collagen. This allows for highly selective isolation and recovery of the cultured animal cells which can be accomplished without incurring any injury from the chemically modified collagen.
Inventors
- 👤 Shunji Kasai
- 👤 Toshihiro Akaike
- 👤 Teruo Miyata
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-23 06:35
CPC Classifications
C; C12; C12N; C12N5/00; C12N5/0068; C12N2533/00; C12N2533/50; C12N2533/54; Y; Y10; Y10S; Y10S435/00; Y10S435/8215; Y10S435/948
Claims Analysis
Total Claims: 13
Claim 1
Independent Claim
42 words
In a mammalian cell culture containing a substratum and nutrient medium, the improvement comprising a substratum composed of collagen which has been modified by chemical means to increase the charge of the collagen to a degree sufficient to enhance mammalian cell attachment.
Claim 2
Dependent Claim
42 words
In a mammalian cell culture containing a substratum and nutrient medium, the improvement comprising a substratum composed of collagen which has been modified by chemical means to increase the charge of the collagen to a degree sufficient to enhance mammalian cell attachment.
Claim 3
Dependent Claim
32 words
The cell culture according to claim 1, wherein the collagen is modified by chemical means to increase the negative charge of the collagen to a degree sufficient to enhance mammalian cell attachment.
Claim 4
Dependent Claim
32 words
The cell culture according to claim 1, wherein the collagen is modified by chemical means to increase the positive charge of the collagen to a degree sufficient to enhance mammalian cell attachment.
Claim 5
Dependent Claim
15 words
The cell culture according to claim 2, wherein the chemically modified collagen is succinylated collagen.
Claim 6
Dependent Claim
15 words
The cell culture according to claim 3, wherein the chemically modified collagen is esterified collagen.
Claim 7
Dependent Claim
23 words
The cell culture according to claim 4, wherein the succinylated collagen is succinylated more than 20 mol% of total amino groups of collagen.
Claim 8
Dependent Claim
23 words
The cell culture according to claim 3, wherein the esterified collagen is esterified more than 20 mol% of total carboxyl groups of collagen.
Claim 9
Dependent Claim
83 words
An improved method for culturing and isolating mammalian cells wherein the cells are cultured under conventional culture conditions, according to the type of cell being cultured, and collagen is used as a substratum for attaching the cells during culturing, the improvement comprising: adhering the mammalian cells to collagen which has been modified by chemical means to increase the charge of the collagen, and incubating the mammalian cells and treating the animal cells with bivalent cation chelating agent to isolate and recover the cells.
Claim 10
Dependent Claim
83 words
An improved method for culturing and isolating mammalian cells wherein the cells are cultured under conventional culture conditions, according to the type of cell being cultured, and collagen is used as a substratum for attaching the cells during culturing, the improvement comprising: adhering the mammalian cells to collagen which has been modified by chemical means to increase the charge of the collagen, and incubating the mammalian cells and treating the animal cells with bivalent cation chelating agent to isolate and recover the cells.
Claim 11
Dependent Claim
24 words
A method according to claim 8, wherein macrophages are incubated in the mixed system with lymphocytes and only macrophages are selectively isolated and recovered.
Claim 12
Dependent Claim
16 words
A method according to claim 8, wherein the bi-valent catin chelating agent is EDTA or EGTA.
Claim 13
Dependent Claim
26 words
A method according to claim 8, wherein the adhesion and incubation of mammalian cells are carried out in the presence of bovine fetus serum or fibronectin.
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) | 1988-12-16 | 1989-06-17 | 1989-12-17 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1992-12-16 | 1993-06-17 | 1993-12-17 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1996-12-16 | 1997-06-17 | 1997-12-17 | $7700.00 | Window Closed / Lapsed |