US4189534A
Expired / Lapsed
US
⏱️ 0.0 years remaining
Cell culture microcarriers
Assignee: Massachusetts Institute of Technology
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Filing: 1977-10-17
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Granted: 1980-02-19
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Expires: 1997-10-17
Abstract
Abstract Improved cell culture microcarriers, and methods for their production and use, are disclosed herein. These improved microcarriers have positive charge capacities adjusted and/or controlled within a range suitable for good cell growth. One method for producing such improved microcarriers is by treating beads formed from polymers containing pendant hydroxy groups, such as dextran beads, with an aqueous solution of an alkaline material and a chloro- or bromo-substituted tertiary amine under precisely controlled conditions to produce the desired exchange capacity. The resultant positively charged microcarriers have been used in microcarrier cultures to produce outstanding growth of anchorage-dependent cells. Such cells can be harvested, or used for the production of viruses, vaccines, hormones, interferon or other cellular growth by-products.
Inventors
- 👤 David W. Levine
- 👤 William G. Thilly
- 👤 Daniel I. C. Wang
- 👤 Jason S. Wong
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-23 19:23
CPC Classifications
C; C12; C12N; C12N5/00; C12N5/0068; C12N5/0075; C07; C07K; C07K14/00; C07K14/435; C07K14/52; C07K14/555; C07K14/565; C12M; C12M25/00; C12M25/16; A; A61; A61K; A61K38/00; C12N2531/00; C12N2533/00; C12N2533/70; Y; Y10; Y10S; Y10S435/00; Y10S435/811
Claims Analysis
Total Claims: 24
Claim 1
Independent Claim
56 words
In a method of growing anchorage-dependent cells in microcarrier culture, the improvement of employing microcarriers having an amount of positively charged chemical moieties thereon adjusted to provide an exchange capacity which allows good growth of said cells, said exchange capacity being within the range of between about 0.1 and about 4.5 meq/gram of dry, untreated microcarriers.
Claim 2
Dependent Claim
56 words
In a method of growing anchorage-dependent cells in microcarrier culture, the improvement of employing microcarriers having an amount of positively charged chemical moieties thereon adjusted to provide an exchange capacity which allows good growth of said cells, said exchange capacity being within the range of between about 0.1 and about 4.5 meq/gram of dry, untreated microcarriers.
Claim 3
Dependent Claim
15 words
In the method of claim 1, the improvement wherein said microcarriers comprise crosslinked dextran beads.
Claim 4
Dependent Claim
25 words
In the method of claim 2, the improvement wherein the positively charged chemical moieties on said crosslinked dextran beads comprise tertiary or quaternary amine groups.
Claim 5
Dependent Claim
29 words
In the method of claim 3, the improvement wherein said exchange capacity is within the range of from about 1 to about 2.8 meq/gram of dry, untreated crosslinked dextran.
Claim 6
Dependent Claim
22 words
In the method of claim 4, the improvement wherein the positively charged chemical moieties on said crosslinked dextran beads comprise diethylaminoethyl groups.
Claim 7
Dependent Claim
24 words
In the method of claim 5, the improvement wherein said dry, crosslinked beads have a diameter of approximately 75 μm in their dry state.
Claim 8
Independent Claim
75 words
A method of growing anchorage-dependent cells, comprising: a. forming a suspension in cell culture medium of positively charged microcarriers having their charge capacity adjusted to a value which supports good growth of cells, said charge capacity being within the range of from about 0.1 to about 4.5 meq/gram of dry, untreated microcarriers; b. inoculating cells into said suspension of microcarriers to form a cell culture; and, c. maintaining said cell culture under cell growth conditions.
Claim 9
Independent Claim
75 words
A method of growing anchorage-dependent cells, comprising: a. forming a suspension in cell culture medium of positively charged microcarriers having their charge capacity adjusted to a value which supports good growth of cells, said charge capacity being within the range of from about 0.1 to about 4.5 meq/gram of dry, untreated microcarriers; b. inoculating cells into said suspension of microcarriers to form a cell culture; and, c. maintaining said cell culture under cell growth conditions.
Claim 10
Dependent Claim
19 words
A method of claim 7 wherein said microcarriers comprise crosslinked dextran beads having tertiary or quaternary amine groups thereon.
Claim 11
Dependent Claim
16 words
A method of claim 7 wherein said microcarriers comprise crosslinked dextran beads having diethylaminoethyl groups thereon.
Claim 12
Dependent Claim
28 words
A method of claim 9 wherein said microcarriers have a charge capacity within the range of between about 1 and about 2.8 meq/gram of dry, untreated crosslinked dextran.
Claim 13
Dependent Claim
16 words
A method of claim 10 wherein said microcarriers have an average diameter of about 75 μm.
Claim 14
Dependent Claim
104 words
In the method of growing anchorage-dependent cells including forming a suspension in cell culture medium of microcarriers comprising crosslinked dextran beads, inoculating cells into said suspension to form a cell culture and maintaining said cell culture under cell growth conditions: The improvement of pre-treating said crosslinked dextran beads by reacting them in an aqueous solution of a tertiary or quaternary amine and a base under conditions sufficient to provide a positive charge capacity on said beads which is suitable for exponential growth of said anchorage-dependent cells, said charge capacity being within the range between about 0.1 and about 4.5 meq/gram of dry, untreated microcarriers.
Claim 15
Dependent Claim
104 words
In the method of growing anchorage-dependent cells including forming a suspension in cell culture medium of microcarriers comprising crosslinked dextran beads, inoculating cells into said suspension to form a cell culture and maintaining said cell culture under cell growth conditions: The improvement of pre-treating said crosslinked dextran beads by reacting them in an aqueous solution of a tertiary or quaternary amine and a base under conditions sufficient to provide a positive charge capacity on said beads which is suitable for exponential growth of said anchorage-dependent cells, said charge capacity being within the range between about 0.1 and about 4.5 meq/gram of dry, untreated microcarriers.
Claim 16
Independent Claim
83 words
A method for producing anchorage-dependent cell growth by-products comprising: a. forming a suspension of positively-charged microcarriers having a charge capacity sufficient for good growth of anchorage dependent cells in a suitable cell culture medium, said charge capacity being between about 0.1 and about 4.5 meq/gram of dry, untreated microcarriers; b. inoculating said culture with anchorage-dependent cells to form a cell culture; c. maintaining said cell culture under conditions conducive to the production of cell growth by-products; and, d. harvesting said cell growth by-products.
Claim 17
Independent Claim
83 words
A method for producing anchorage-dependent cell growth by-products comprising: a. forming a suspension of positively-charged microcarriers having a charge capacity sufficient for good growth of anchorage dependent cells in a suitable cell culture medium, said charge capacity being between about 0.1 and about 4.5 meq/gram of dry, untreated microcarriers; b. inoculating said culture with anchorage-dependent cells to form a cell culture; c. maintaining said cell culture under conditions conducive to the production of cell growth by-products; and, d. harvesting said cell growth by-products.
Claim 18
Dependent Claim
13 words
A method of claim 13 wherein said cell growth by-product is a virus.
Claim 19
Dependent Claim
13 words
A method of claim 13 wherein said cell growth by-product is a hormone.
Claim 20
Dependent Claim
12 words
A method of claim 13 wherein said cell growth by-product is interferon.
Claim 21
Dependent Claim
29 words
A method of claim 13 wherein said microcarriers comprise a reaction product of crosslinked dextran beads and an aqueous solution of a tertiary or quaternary amine and a base.
Claim 22
Dependent Claim
16 words
A method of claim 13 wherein said microcarriers comprise crosslinked dextran beads having diethylaminoethyl groups thereon.
Claim 23
Dependent Claim
25 words
A method of claim 18 wherein said charge capacity is in the range of from about 1 to about 2.8 meq/grams of dry, untreated microcarriers.
Claim 24
Dependent Claim
20 words
A method of claim 19 wherein said reaction product comprises hydrated beads having an average diameter of about 120-200 μm.
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) | 1983-02-18 | 1983-08-20 | 1984-02-19 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1987-02-18 | 1987-08-20 | 1988-02-19 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1991-02-18 | 1991-08-20 | 1992-02-19 | $7700.00 | Window Closed / Lapsed |