US3887430A
Expired / Lapsed
US
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Culture medium for tissue culture techniques
Assignee: Dow Chemical Co
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Filing: 1973-07-09
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Granted: 1975-06-03
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Expires: 1993-07-09
Abstract
Abstract A chemically defined, protein-free culture medium embodying an anion-exchange resin and a water-soluble lipid source provides means for tissue culture.
Inventors
- 👤 Harry L Torney
- 👤 Helen T Torney
- 👤 Dale E Bordt
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-23 18:32
CPC Classifications
C; C12; C12N; C12N5/00; C12N5/0018; C12N5/005; C12N2500/00; C12N2500/30; C12N2500/36; C12N2500/50
Claims Analysis
Total Claims: 5
Claim 1
Independent Claim
178 words
IN A TISSUE CULTURE METHOD COMPRISING THE STEPS OF INOCULATING A PROTEIN-FREE, STERILE TISSUE CULTURE MEDIUM WITH LIVING ANIMAL TISSUE CELLS AND INCUBATING SAID INOCULATED TISSUE CULTURE MEDIUM UNDER CONDITIONS CONDUCTIVE TO CELL PROPAGATION, THE IMPROVEMENT WHICH COMPRISES THE STEPS OF MIXING A GROWTH-ENHANCING AMOUNT OF A WATER-SOLUBLE LIPID SOURCE SELECTED FROM THE GROUP CONSISTING OF POLYSORBATE 60 AND POLYSORBATE 80 WITH SAID MEDIUM, IN AN AMOUNT OF FROM ABOUT 0.0010 TO ABOUT 0.100 PART BY VOLUME OF LIPID SOURCE PER 100 PARTS BY VOLUME OF MEDIUM, INTIMATELY CONTACTING SAID MEDIUM WITH A GROWTH-ENHANCING AMOUNT OF A STRONGLY BASIC ANION-EXHANCE RESIN OBTAINED BY REACTING TRIMETHYLAMINE WITH A HALOMETHYLATED POLYMER PREPARED BY REACTING CHLOROMETHYL METHYL ETHER IN THE PRESENCE OF A ZINC CHLORIDE CATALYST WITH BEADS OF A COPOLYMER OF ABOUT 98 PERCENT OF STYRENE AND 2 PERCENT OF DIVINYLBENZENE AND THEREAFTER INCUBATING SAID TIS SUE CULTURE MEDIUM, CELLS AND POLYSORBATE LIPID SOURCE IN INTIMATE CONTACT WITH SAID STRONGLY BASIC ANION-EXCHANGE RESIN UNDER CONDITIONS CONDUCTIVE TO CELL PROPAGATION FOR A TIME SUFFICIENT TO OBTAIN SUBSTANTIAL PROGATION OF SAID CELLS.
Claim 2
Dependent Claim
178 words
IN A TISSUE CULTURE METHOD COMPRISING THE STEPS OF INOCULATING A PROTEIN-FREE, STERILE TISSUE CULTURE MEDIUM WITH LIVING ANIMAL TISSUE CELLS AND INCUBATING SAID INOCULATED TISSUE CULTURE MEDIUM UNDER CONDITIONS CONDUCTIVE TO CELL PROPAGATION, THE IMPROVEMENT WHICH COMPRISES THE STEPS OF MIXING A GROWTH-ENHANCING AMOUNT OF A WATER-SOLUBLE LIPID SOURCE SELECTED FROM THE GROUP CONSISTING OF POLYSORBATE 60 AND POLYSORBATE 80 WITH SAID MEDIUM, IN AN AMOUNT OF FROM ABOUT 0.0010 TO ABOUT 0.100 PART BY VOLUME OF LIPID SOURCE PER 100 PARTS BY VOLUME OF MEDIUM, INTIMATELY CONTACTING SAID MEDIUM WITH A GROWTH-ENHANCING AMOUNT OF A STRONGLY BASIC ANION-EXHANCE RESIN OBTAINED BY REACTING TRIMETHYLAMINE WITH A HALOMETHYLATED POLYMER PREPARED BY REACTING CHLOROMETHYL METHYL ETHER IN THE PRESENCE OF A ZINC CHLORIDE CATALYST WITH BEADS OF A COPOLYMER OF ABOUT 98 PERCENT OF STYRENE AND 2 PERCENT OF DIVINYLBENZENE AND THEREAFTER INCUBATING SAID TIS SUE CULTURE MEDIUM, CELLS AND POLYSORBATE LIPID SOURCE IN INTIMATE CONTACT WITH SAID STRONGLY BASIC ANION-EXCHANGE RESIN UNDER CONDITIONS CONDUCTIVE TO CELL PROPAGATION FOR A TIME SUFFICIENT TO OBTAIN SUBSTANTIAL PROGATION OF SAID CELLS.
Claim 3
Dependent Claim
177 words
In a tissue culture method comprising the steps of inoculating a protein-free, sterile tissue culture medium with living animal tissue cells and incubating said inoculated tissue culture medium under conditions conductive to cell propagation, the improvement which comprises the steps of mixing a growth-enhancing amount of a water-soluble lipid source selected from the group consisting of polysorbate 60 and polysorbate 80 with said medium, in an amount of from about 0.0010 to about 0.100 part by volume of lipid source per 100 parts by volume of medium, intimately contacting said medium with a growth-enhancing amount of a strongly basic anion-exhance resin obtained by reacting trimethylamine with a halomethylated polymer prepared by reacting chloromethyl methyl ether in the presence of a zinc chloride catalyst with beads of a copolymer of about 98 percent of styrene and 2 percent of divinylbenzene and thereafter incubating said tissue culture medium, cells and polysorbate lipid source in intimate contact with said strongly basic anion-exchange resin under conditions conducive to cell propagation for a time sufficient to obtain substantial propagation of said cells.
Claim 4
Dependent Claim
177 words
In a tissue culture method comprising the steps of inoculating a protein-free, sterile tissue culture medium with living animal tissue cells and incubating said inoculated tissue culture medium under conditions conductive to cell propagation, the improvement which comprises the steps of mixing a growth-enhancing amount of a water-soluble lipid source selected from the group consisting of polysorbate 60 and polysorbate 80 with said medium, in an amount of from about 0.0010 to about 0.100 part by volume of lipid source per 100 parts by volume of medium, intimately contacting said medium with a growth-enhancing amount of a strongly basic anion-exhance resin obtained by reacting trimethylamine with a halomethylated polymer prepared by reacting chloromethyl methyl ether in the presence of a zinc chloride catalyst with beads of a copolymer of about 98 percent of styrene and 2 percent of divinylbenzene and thereafter incubating said tissue culture medium, cells and polysorbate lipid source in intimate contact with said strongly basic anion-exchange resin under conditions conducive to cell propagation for a time sufficient to obtain substantial propagation of said cells.
Claim 5
Dependent Claim
13 words
The method of claim 1 wherein the lipid source is polysorbate 60. 80.
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) | 1978-06-02 | 1978-12-02 | 1979-06-03 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1982-06-02 | 1982-12-02 | 1983-06-03 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1986-06-02 | 1986-12-02 | 1987-06-03 | $7700.00 | Window Closed / Lapsed |