Bing [CAK 8, CAK8] (ATCC® CRL-11554)

Organism: Homo sapiens, human  /  Cell Type: epithelialtransformed with adenovirus 5 DNA  / 

Permits and Restrictions

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Organism Homo sapiens, human
Cell Type epithelialtransformed with adenovirus 5 DNA
Product Format frozen
Morphology epithelial
Culture Properties adherent
Biosafety Level 2 Cells contain adenovirus 5 DNA sequences
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
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
Protocol:
  1. Remove and discard culture medium.
  2. 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.
  3. 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.
  4. Add 6.0 to 8.0 ml of complete growth medium and aspirate cells by gently pipetting.
  5. Add appropriate aliquots of the cell suspension to new culture vessels.
  6. Incubate cultures at 37°C.
Subcultivation Ratio: 1:4 to 1:6
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
Atmosphere: air, 95%; carbon dioxide (CO2), 5%
Temperature: 37.0°C
Name of Depositor Rockefeller Univ.
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.

Basic Documentation
Restrictions

The line is available with the following restriction: 1. The cell line was deposited at the ATCC by Rockefeller University and is provided for research purposes only. Neither the cell line nor the products derived from it may be sold or used for commercial purposes. Nor can the cells be distributed to third parties for purposes of sale, or producing for sale, cells or their products. The cells are provided as a service to the research community. They are provided without warranty of merchantability or fitness for a particular purpose or any other warranty, expressed or implied. 2.Any proposed commercial use of the cells, or their products, must first be negotiated with Rockefeller University, Office of Technology Transfer, 1230 York Avenue, New York, NY 10065 Attn: Kathleen A. Denis, Associate Vice President Technology Transfer.

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.