PA317 LXSN 16E7 (ATCC® CRL-2205)

Organism: Mus musculus, mouse  /  Cell Type: Retinoblastoma,keratinocyte  /  Disease: Papilloma,Retinoblastoma

Permits and Restrictions

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Organism Mus musculus, mouse
Cell Type Retinoblastoma,keratinocyte
Product Format frozen
Morphology fibroblast
Culture Properties adherent
Biosafety Level 2 Cells contain human papilloma viral DNA sequences
Disease Papilloma,Retinoblastoma
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 pLXSN16E7 vector contains the human papilloma virus (HPV) type 16 E7 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 LXSN16E6 which encodes the HPV16 E7 open reading frame, and which can be used to stably infect many cell types.
The resulting cells constitutively express the E6 protein of HPV16 which binds to and inactivates the retinoblastoma (Rb) gene.
The virus can be used to immortalize human keratinocytes.
Derivation
PA317 LXSN 16E7 is a packaging cell line developed by transfection of the retrovirus vector pLXSN16E7 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 16E7 is a packaging cell line developed by transfection of the retrovirus vector pLXSN16E7 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 pLXSN16E7 vector contains the human papilloma virus (HPV) type 16 E7 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 LXSN16E6 which encodes the HPV16 E7 open reading frame, and which can be used to stably infect many cell types.
The resulting cells constitutively express the E6 protein of HPV16 which binds to and inactivates the retinoblastoma (Rb) gene.
The virus can be used to immortalize human keratinocytes.
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 16E7 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