| 產品名稱 |
Phoenix-AMPHO |
| 商品貨號 |
B161447 |
| Organism |
Homo sapiens, human |
| Tissue |
kidney |
| Product Format |
frozen |
| Morphology |
epithelial |
| Culture Properties |
adherent |
| Biosafety Level |
2 [Cells contain Adeno and SV-40 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. |
| Age |
embryo |
| Applications |
Phoenix is a second-generation retrovirus producer cell line for the generation of helper-free ecotropic and amphotropic retroviruses.
The unique feature of this cell line is that it is highly transfectable with either calcium phosphate mediated transfection or lipid-based transfection protocols-- up to 50% or higher of cells can be transiently transfected.
This cell line is capable of carrying episomes for long-term stable production of retrovirus (LZRS). |
| Storage Conditions |
liquid nitrogen vapor phase |
| Derivation |
HEK 293T/17 cells were transformed with adenovirus E1a carrying a temperature sensitive T antigen co-selected with neomycin. Transformation was brought about by the insertion of approximately 4.5 kilobases of viral genome into human chromosome 19. |
| Comments |
Phoenix Amphotropic (AMPHO) cell line expresses amphotropic envelope protein.
CD8 expression is a direct reflection of intracellular gag-pol and the stability of the producer cell population's ability to produce gag-pol can be readily monitored by flow cytometry.
Phoenix-Ampho have been extensively tested for helper virus production and established as being helper-virus free.
|
| Complete Growth Medium |
The base medium for this cell line is Dulbecco’s Modified Eagle’s Medium (DMEM) (ATCC 30-2002). To make the complete growth medium, add the following components to the base medium: 10% Fetal Bovine Serum (heat inactivated) (ATCC 30-2020), 2mM L-glutamine (ATCC 30-2214), 1% Penicillin/Streptomycin.
|
| Subculturing |
Volumes are given for a 75 cm2 flask. Increase or decrease the amount of dissociation medium needed proportionally 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.
Note: Culture to ≤80% confluence to prevent the cells from sloughing
Subcultivation Ratio: A subcultivation ratio of 1:5 to 1:8 is recommended
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 |
Temperature: 37°C
Atmosphere: air, 95%; carbon dioxide (CO2), 5% |
| Cells per Vial |
4.5 x 106 cells per vial |
| STR Profile |
Amelogenin: X
CSF1PO: 11,12
D13S317: 11,12,13,14
D16S539: 9,13
D5S818: 8,10
D7S820: 11
TH01: 7,9.3
TPOX: 11
vWA: 16,18,19
|
| Population Doubling Time |
16 hours |
| Name of Depositor |
G Nolan |
| Year of Origin |
1996 |
| References |
Swift S et al. Rapid Production of Retroviruses for Efficient Gene Delivery to Mammalian Cells Using 293T Cell-Based Systems. Current Protocols in Immunology,Unit 10.17C, 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
Nolan Laboratory Retroviral Systems. Phoenix helper-free retrovirus producer lines, https://web.stanford.edu/group/nolan/_OldWebsite/retroviral_systems/retsys.html
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