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微信小章| 产品名称： | Bing CAK 8, CAK8 |
|---|---|
| 商品货号： | TS215176 |
| Organism： | Homo sapiens, human |
| Tissue： | kidney |
| Cell Type： | epithelialtransformed with adenovirus 5 DNA |
| Product Format： | frozen |
| Morphology： | epithelial |
| Culture Properties： | adherent |
| Biosafety Level： | 2 Cells contain adenovirus 5 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： | Individual clones were isolated and tested for the ability to produce high titer beta galactosidase expressing retroviruses. The amphotropic envelope expressing construct, pCripAMgag- which contains mutations in the gag region, lacks the packaging site, and replaces the 3 LTR, was transfected into Anjou 65 cells along with a plasmid expressing the gp resistance gene. Bing is an amphotropic envelope-expressing packaging line derived from the 293T cell line. |
| 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： | Bing is an amphotropic envelope-expressing packaging line derived from the 293T cell line. 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 (see ATCC CRL-11268). 293T/17 cells were cotransfected with the pCRIPenv- and the pCRIPgag-2 vectors to obtain the ANJOU 65 (see ATCC CRL-11269) cell line. The amphotropic envelope expressing construct, pCripAMgag- which contains mutations in the gag region, lacks the packaging site, and replaces the 3 LTR, was transfected into Anjou 65 cells along with a plasmid expressing the gp resistance gene. Individual clones were isolated and tested for the ability to produce high titer beta galactosidase expressing retroviruses. One clone produced betagal retrovirus with a titer in excess of 10(6)/ml following transfection with pBND8. Two rounds of limiting dilution subcloning were performed subsequently, giving rise to the CAK8, or Bing cell line. |
| Comments： | Bing is an amphotropic envelope-expressing packaging line derived from the 293T cell line. 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 (see ATCC CRL-11268). 293T/17 cells were cotransfected with the pCRIPenv- and the pCRIPgag-2 vectors to obtain the ANJOU 65 (see ATCC CRL-11269) cell line. The amphotropic envelope expressing construct, pCripAMgag- which contains mutations in the gag region, lacks the packaging site, and replaces the 3 LTR, was transfected into Anjou 65 cells along with a plasmid expressing the gp resistance gene. Individual clones were isolated and tested for the ability to produce high titer beta galactosidase expressing retroviruses. One clone produced betagal retrovirus with a titer in excess of 10(6)/ml following transfection with pBND8. Two rounds of limiting dilution subcloning were performed subsequently, giving rise to the CAK8, or Bing cell line. |
| Complete Growth Medium： | Dulbeccos Modified Eagles Medium with 4 mM L-glutamine that is modified by ATCC to contain 4.5 g/L glucose and 1.5 g/L sodium bicarbonate and supplemented with an additional 2 mM L-glutamine, 2176 ng/ml Aminopterin, 0.00978 mg/ml Thymidine, 0.0136 mg/ml Hypoxanthine, 0.025 mg/ml Mycophenolic acid, 0.250 mg/ml Xanthine and 10% dialyzed fetal bovine serum
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| Subculturing： | Volumes used in this protocol are for 75 cm2 flask; proportionally reduce or increase amount of dissociation medium for culture vessels of other sizes.
Subcultivation Ratio: 1:4 to 1:6 Note: For more information on enzymatic dissociation and subculturing of cell lines consult Chapter 10 in Culture of Animal Cells, a Manual of Basic Technique by R. Ian Freshney, 3rd edition, published by Alan R. Liss, N.Y., 1994. |
| Cryopreservation： | Complete growth medium described above supplemented with 5% (v/v) DMSO.xa0 Cell culture tested DMSO is available as ATCC Catalog No. 4-X. |
| Culture Conditions： | Temperature: 37°C
Atmosphere: Air, 95%; Carbon dioxide (CO2), 5% |
| Name of Depositor： | Rockefeller Univ. |
| Deposited As： | human |
| U.S. Patent Number： | |
| References： | 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 Bing is an amphotropic envelope-expressing packaging line derived from the 293T cell line. 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 (see ATCC CRL-11268). 293T/17 cells were cotransfected with the pCRIPenv- and the pCRIPgag-2 vectors to obtain the ANJOU 65 (see ATCC CRL-11269) cell line. The amphotropic envelope expressing construct, pCripAMgag- which contains mutations in the gag region, lacks the packaging site, and replaces the 3 LTR, was transfected into Anjou 65 cells along with a plasmid expressing the gp resistance gene. Individual clones were isolated and tested for the ability to produce high titer beta galactosidase expressing retroviruses. One clone produced betagal retrovirus with a titer in excess of 10(6)/ml following transfection with pBND8. Two rounds of limiting dilution subcloning were performed subsequently, giving rise to the CAK8, or Bing cell line. Hay, R. J., Caputo, J. L., and Macy, M. L., Eds. (1992), ATCC Quality Control Methods for Cell Lines. 2nd edition, Published by ATCC. Caputo, J. L., Biosafety procedures in cell culture. J. Tissue Culture Methods 11:223-227, 1988. Biosafety in Microbiological and Biomedical Laboratories, 5th ed. HHS. U.S. Department of Health and Human Services, Centers for Disease Control and Prevention. Washington DC: U.S. Government Printing Office; 2007. The entire text is available online. |