| 產(chǎn)品名稱 |
293T/17 [HEK 293T/17] |
| 商品貨號(hào) |
B161450 |
| Organism |
Homo sapiens, human |
| Tissue |
kidney |
| Product Format |
frozen |
| Morphology |
epithelial |
| Culture Properties |
adherent |
| Biosafety Level |
2 [Cells contain Adeno and SV-40 viral DNA sequences]
Biosafety classification is based on U.S. Public Health Service Guidelines, it is the responsibility of the customer to ensure that their facilities comply with biosafety regulations for their own country. |
| Age |
fetus |
| Applications |
These cells constitutively express the simian virus 40 (SV40) large T antigen, and clone 17 was selected specifically for its high transfectability. |
| Storage Conditions |
liquid nitrogen vapor phase |
| Disclosure |
This material is cited in a US or other Patent and may not be used to
infringe the claims. Depending on the wishes of the Depositor, ATCC may be
required to inform the Patent Depositor of the party to which the material was
furnished. This material may not have been produced or characterized by ATCC. |
| Derivation |
The 293T/17 cell line is a derivative of the 293T (293tsA1609neo) cell line. 293T is a highly transfectable derivative of the 293 cell line into which the temperature sensitive gene for SV40 T-antigen was inserted. 293T cells were cloned and the clones tested with the pBND and pZAP vectors to obtain a line capable of producing high titers of infectious retrovirus, 293T/17. |
| Antigen Expression |
SV40 T antigen |
| Genes Expressed |
SV40 T antigen |
| Comments |
293T/17 cells were cotransfected with the pCRIPenv- and the pCRIPgag-2 vectors to obtain the ANJOU 65 (see ATCC CRL-11269) cell line. ANJOU 65 cells were cotransfected with the pCRIPgag-2 and pGPT2E vectors to obtain the BOSC 23 (see ATCC CRL-11270) ecotropic envelope-expression packaging cell line. ANJOU 65 cells were also cotransfected with the pCRIPAMgag vector along with a plasmid expressing the gpt resistance gene to obtain the Bing (see ATCC CRL-11554) amphotropic envelope-expression packaging cell line. |
| Complete Growth Medium |
The base medium for this cell line is ATCC-formulated Dulbecco's Modified Eagle's Medium, Catalog No. 30-2002. To make the complete growth medium, add the following components to the base medium: fetal bovine serum to a final concentration of 10%.
|
| Subculturing |
Volumes are given for a 75 cm 2 flask. Increase or decrease the amount of dissociation medium needed proportionally for culture vessels of other sizes. Corning ® T-75 flasks (catalog #430641) are recommended for subculturing this product.
- Remove and discard culture medium.
-
Briefly rinse the cell layer with 0.25% (w/v) Trypsin- 0.53 mM EDTA solution to remove all traces of serum that contains trypsin inhibitor.
-
Add 2.0 to 3.0 mL of Trypsin-EDTA solution to flask and observe cells under an inverted microscope until cell layer is dispersed (usually within 5 to 15 minutes).
Note: To avoid clumping do not agitate the cells by hitting or shaking the flask while waiting for the cells to detach. Cells that are difficult to detach may be placed at 37°C to facilitate dispersal.
-
Add 6.0 to 8.0 mL of complete growth medium and aspirate cells by gently pipetting.
-
Add appropriate aliquots of the cell suspension to new culture vessels.
- Incubate cultures at 37°C.
Subcultivation Ratio: A subcultivation ratio of 1:4 to 1:8 is recommended
Medium Renewal: Every 2 to 3 days |
| Cryopreservation |
Freeze medium: Complete growth medium supplemented with 5% (v/v) DMSO Storage temperature: liquid nitrogen vapor phase |
| Culture Conditions |
Temperature: 37°C
Atmosphere: air, 95%; carbon dioxide (CO2), 5% |
| STR Profile |
Amelogenin: X CSF1PO: 11, 12 D13S317: 12, 14 D16S539: 9, 13 D5S818: 8, 9 D7S820: 11 THO1: 7, 9.3 TPOX: 11 vWA: 16, 18, 19 |
| Name of Depositor |
Rockefeller Univ. |
| Deposited As |
human |
| U.S. Patent Number |
|
| References |
Sena-Esteves M, et al. Single-step conversion of cells to retrovirus vector producers with herpes simplex virus-Epstein-Barr virus hybrid amplicons. J. Virol. 73: 10426-10439, 1999. PubMed: 10559361
Pensiero M, et al. Retroviral vectors produced by producer cell lines resistant to lysis by human serum. US Patent 5,952,225 dated Sep 14 1999
Pensiero M, et al. Retroviral vectors produced by producer cell lines resistant to lysis by human serum. US Patent 6,329,199 dated Dec 11 2001
Pear WS, et al. Production of High-Titer Helper-Free Retroviruses by Transient Transfection. Proc. Natl. Acad. Sci. USA 90: 8392-8396, 1993. PubMed: 7690960
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