ANJOU 65 (ATCC® CRL-11269)

Organism: Homo sapiens, human  / 

Permits and Restrictions

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Organism Homo sapiens, human
Product Format frozen
Morphology epithelial
Culture Properties adherent
Biosafety Level 2 Cells contain SV-40 viral DNA sequences
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 phaseliquid nitrogen vapor phase
Derivation
The ANJOU 65 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). These cells constitutively express the simian virus 40 (SV40) large T antigen, and clone 17 was selected specifically for its high transfectability. 293T/17 cells were cotransfected with the pCRIPenv- and the pCRIPgag-2 vectors to obtain the ANJOU 65 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.
Comments
The ANJOU 65 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). These cells constitutively express the simian virus 40 (SV40) large T antigen, and clone 17 was selected specifically for its high transfectability. 293T/17 cells were cotransfected with the pCRIPenv- and the pCRIPgag-2 vectors to obtain the ANJOU 65 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
Protocol: Remove medium, and rinse with 0.25% trypsin, 0.03% EDTA solution. Remove the solution and add an additional 1 to 2 ml of trypsin-EDTA solution. Allow the flask to sit at room temperature (or at 37C) until the cells detach. Add fresh culture medium, aspirate and dispense into new culture flasks. Do not let monolayer get too heavy or cells will slough off.
Subcultivation Ratio: A subcultivation ratio of 1:4 to 1:8 is recommended
Cryopreservation
Freeze medium: Complete growth medium 95%; DMSO, 5%
Storage temperature: liquid nitrogen vapor phaseliquid nitrogen vapor phase
Culture Conditions
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

Notice: Necessary PermitsPermits

These permits may be required for shipping this product:

  • Customers located in the state of Hawaii will need to contact the Hawaii Department of Agriculture to determine if an Import Permit is required. A copy of the permit or documentation that a permit is not required must be sent to ATCC in advance of shipment.
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