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Bing [CAK 8, CAK8]
Bing [CAK 8, CAK8]
規(guī)格:
貨期:
編號:B161448
品牌:Mingzhoubio

標(biāo)準(zhǔn)菌株
定量菌液
DNA
RNA

規(guī)格:
凍干粉
斜面
甘油
平板


產(chǎn)品名稱 Bing [CAK 8, CAK8]
商品貨號 B161448
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 Dulbecco's Modified Eagle's 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
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.
  1. Remove and discard culture medium.
  2. 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.
  3. Add 6.0 to 8.0 mL of complete growth medium and aspirate cells by gently pipetting.
  4. Add appropriate aliquots of the cell suspension to new culture vessels.
  5. Incubate cultures at 37°C.

Subcultivation Ratio: 1:4 to 1:6
Medium Renewal: Every 2 to 3 days

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.  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.

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