PA317 LXSN 6E6 (ATCC® CRL-2206)

Organism: Mus musculus, mouse  /  Disease: Papilloma

Permits and Restrictions

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Organism Mus musculus, mouse
Product Format frozen
Morphology fibroblast
Culture Properties adherent
Biosafety Level 2 Cells contain human papilloma viral DNA sequences
Disease Papilloma
Age embryo
Strain NIH/Swiss
Applications
Virions produced from the transfected Psi-2 cells were used to infect the amphotropic packaging line PA317, and infected cells were selected in medium containing G418.
The pLXSN6bE6 vector contains the human papilloma virus (HPV) type 6b E6 gene under control of the Moloney murine leukemia virus (MoMuLV) promoter - enhancer sequences.
The vector also contains a gene controlling resistance to neomycin transcribed from the SV40 promoter.
This line produces the amphotropic retrovirus LXSN6bE6 which encodes the HPV6b E6 open reading frame, and which can be used to stably infect many cell types.
Derivation
PA317 LXSN 6E6 is a packaging cell line developed by transfection of the retrovirus vector pLXSN6bE6 into the Psi-2 ecotropic packaging cell line.
Virions produced from the transfected Psi-2 cells were used to infect the amphotropic packaging line PA317, and infected cells were selected in medium containing G418.
Comments
PA317 LXSN 6E6 is a packaging cell line developed by transfection of the retrovirus vector pLXSN6bE6 into the Psi-2 ecotropic packaging cell line.
Virions produced from the transfected Psi-2 cells were used to infect the amphotropic packaging line PA317, and infected cells were selected in medium containing G418.
The pLXSN6bE6 vector contains the human papilloma virus (HPV) type 6b E6 gene under control of the Moloney murine leukemia virus (MoMuLV) promoter - enhancer sequences.
The vector also contains a gene controlling resistance to neomycin transcribed from the SV40 promoter.
This line produces the amphotropic retrovirus LXSN6bE6 which encodes the HPV6b E6 open reading frame, and which can be used to stably infect many cell types.
The resulting cells constitutively express the E6 protein of HPV6b.
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
Subcultivation Ratio: A subcultivation ratio of 1:6 to 1:12 is recommended
Medium Renewal: Every 2 to 3 days
Remove medium, rinse flask with fresh 0.25% trypsin, 0.02% EDTA and remove trypsin, Add an additional 1 to 2 ml of trypsin solution, and allow the flask to sit at room temperature (or 37C) until the cells detach.
Add fresh medium, aspirate and dispense into new flasks.
Cryopreservation
Culture medium, 95%; DMSO, 5%
Name of Depositor DA Galloway
References

Halbert CL, et al. The E7 gene of human papillomavirus type 16 is sufficient for immortalization of human epithelial cells. J. Virol. 65: 473-478, 1991. PubMed: 1845902

Basic Documentation
Restrictions

Note: The user of the PA317 LXSN 6E6 cell line agrees to indemnify and hold harmless the United States, the ATCC, Denise A. Galloway and Fred Hutchinson Cancer Research Center, Seattle, Washington from any claims, costs, damages, or expenses resulting from any injury (including death), damage or loss that may arise from the use of the cell line either directly (including use for diagnostic purposes) or in the preparation of a product. The user assumes all risks and responsibilities in connection with the receipt, handling, storage and use of the material.

References

Halbert CL, et al. The E7 gene of human papillomavirus type 16 is sufficient for immortalization of human epithelial cells. J. Virol. 65: 473-478, 1991. PubMed: 1845902