US4465769A
Expired / Lapsed
US
⏱️ 0.0 years remaining
Non-transformed thymidine kinaseless cell line and its use for testing tumorigenic potential of genes
Assignee: HEALTH AND HUMAN SERVICES United States, AS REPRESENTED BY DEPARTMENT OF
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Filing: 1981-12-11
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Granted: 1984-08-14
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Expires: 2001-12-11
Abstract
Abstract A non-transformed cell line is produced by treating BALB/c derived 10E2 cells with 5-bromodeoxyuridine and 5-iodo-2'-deoxyuridine to produce thymidine kinaseless cells which upon multiple cloning show a flat epitheloid appearance indicative of their non-transforming potential. These cells are used to determine the tumorigenic transforming potential of any gene by introducing the gene into the cells of the non-transformed cell line along with the Herpes simplex virus thymidine kinase gene which serves as a vehicle for cotransfection. The transformation of the cells is indicative of tumorigenic potential.
Inventors
- 👤 Berge Hampar
- 👤 Stephen D. Showalter
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-26 04:18
CPC Classifications
C; C12; C12Q; C12Q1/00; C12Q1/68; Y; Y10; Y10S; Y10S435/00; Y10S435/8215; Y10S435/948
Claims Analysis
Total Claims: 12
Claim 1
Independent Claim
72 words
A method for the determination of the tumorigenic or morphological transforming potential of a non-thymidine kinase (TK) gene comprising, introducing a non-TK gene and a TK gene into an animal cell line which lacks TK and is non-tumorigenic and not transformed morphologically by co-transformation or transformation of ligated genes, selecting for TK containing cells and, testing the resulting cells for tumorigenic potential in animals or phenotypic expression of morphological transformation in vitro.
Claim 2
Independent Claim
72 words
A method for the determination of the tumorigenic or morphological transforming potential of a non-thymidine kinase (TK) gene comprising, introducing a non-TK gene and a TK gene into an animal cell line which lacks TK and is non-tumorigenic and not transformed morphologically by co-transformation or transformation of ligated genes, selecting for TK containing cells and, testing the resulting cells for tumorigenic potential in animals or phenotypic expression of morphological transformation in vitro.
Claim 3
Dependent Claim
21 words
A method according to claim 1 where the cell line lacking TK has all the identifying characteristics of ATCC CRL 8085.
Claim 4
Dependent Claim
56 words
A method according to claim 1 where the cell line lacking TK is produced by the process of exposing TK containing cells to at least one pyrimidine analogue capable of selecting for cells which lack TK activity, selecting and isolating a morpholigically non-transformed, non-tumorigenic animal cell line lacking TK and, cloning the resulting isolated cell line.
Claim 5
Independent Claim
52 words
A method for preparing morphologically non-transformed, non-tumorigenic animal cells which lack thymidine kinase (TK) comprising, exposing TK containing cells to at least one pyrimidine analogue capable of selecting for cells which lack TK activity, selecting and isolating a morphologically non-transformed, non-tumorigenic animal cell line lacking TK and, cloning said isolated cell line.
Claim 6
Independent Claim
52 words
A method for preparing morphologically non-transformed, non-tumorigenic animal cells which lack thymidine kinase (TK) comprising, exposing TK containing cells to at least one pyrimidine analogue capable of selecting for cells which lack TK activity, selecting and isolating a morphologically non-transformed, non-tumorigenic animal cell line lacking TK and, cloning said isolated cell line.
Claim 7
Dependent Claim
14 words
A method according to claim 4 where the pyrimidine analogues are 5-bromodeoxyuridine and 5-iodo-2-deoxyuridine.
Claim 8
Dependent Claim
17 words
A method according to claim 5 where the cells are exposed to 5-bromodeoxyuridine before exposure to 5-iodo-2-deoxyuridine.
Claim 9
Dependent Claim
17 words
A method according to claim 4 where the concentration of pyrimidine analogue is gradually increased during exposure.
Claim 10
Dependent Claim
14 words
A cell line produced by the process of claim 4, 5, 6 or 7.
Claim 11
Dependent Claim
17 words
A non-transformed, non-tumorigenic thymidine kinase lacking cell line having all the identifying characteristics of ATCC CRL 8085.
Claim 12
Dependent Claim
17 words
A non-transformed, non-tumorigenic thymidine kinase lacking cell line having all the identifying characteristics of ATCC CRL 8085.
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) | 1987-08-14 | 1988-02-13 | 1988-08-14 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1991-08-14 | 1992-02-13 | 1992-08-14 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1995-08-14 | 1996-02-13 | 1996-08-14 | $7700.00 | Window Closed / Lapsed |