| 產(chǎn)品名稱 |
PA317 LXSN 16E6E7 |
| 商品貨號 |
B165472 |
| Organism |
Mus musculus, mouse |
| Tissue |
embryo |
| Product Format |
frozen |
| Morphology |
fibroblast |
| Culture Properties |
adherent, adherent |
| Biosafety Level |
2 [Cells contain human papilloma viral 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. |
| Disease |
Papilloma |
| Age |
embryo |
| Strain |
NIH/Swiss |
| Applications |
This cell line produces the amphotropic retrovirus LXSN16E6E7 which encodes the HPV16 E6 and E7 open reading frames, and which can be used to stably infect and immortalize many cell types.
|
| Storage Conditions |
liquid nitrogen vapor phase |
| Derivation |
PA317 LXSN 16E6E7 is a packaging cell line developed by transfection of the retrovirus vector pLXSN16E6E7 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 |
The pLXSN16E6E7 vector contains the human papilloma virus (HPV) type 16 E6 and E7 genes 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.
|
| 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 |
Volumes used in this protocol are for 75 cm2 flask; proportionally reduce or increase amount of dissociation medium for culture vessels of other sizes.
- Remove and discard culture medium.
- 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.
- 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.
- Add 6.0 to 8.0 mL of complete growth medium and aspirate cells by gently pipetting.
- Add appropriate aliquots of the cell suspension to new culture vessels.
- Incubate cultures at 37°C
Subculture Ratio: 1:6 to 1:12
Medium Renewal: 2 to 3 times a week.
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 |
Freeze medium: Complete growth medium supplemented with 5% (v/v) DMSO Storage temperature: liquid nitrogen vapor phase |
| Culture Conditions |
Temperature: 37°C |
| Name of Depositor |
DA Galloway |
| Deposited As |
Mus musculus |
| 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
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