| 產品名稱 |
HeLaRC32 [HeRC32] |
| 商品貨號 |
B197800 |
| Organism |
Homo sapiens, human |
| Tissue |
cervix |
| Product Format |
frozen |
| Morphology |
epithelial |
| Culture Properties |
adherent |
| Biosafety Level |
2 [Cells contain human papilloma virus 18 (HPV-18)]
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 |
adenocarcinoma |
| Age |
31 |
| Gender |
female |
| Ethnicity |
Black |
| Applications |
These cells generate rAAV preparations with high titers of infectious particles which are essentially free of adenoviruses for biological, preclinical, clinical or pharmaceutical applications.
HelaRC32 is a stable packaging cell line expressing the rep and cap genes for recombinant adeno-associated virus type 2 (rAAV-2) assembly which constitutes an attractive alternative to transient transfection protocols. Ref Tessier J, et al. Characterization of adenovirus-induced inverted terminal repeat-independent amplification of integrated adeno-associated virus rep-cap sequences. J. Virol. 75(1):375-383, 2001. PubMed: 11119606 |
| Storage Conditions |
liquid nitrogen vapor phase |
| Images |
 |
| Derivation |
This is a clone of HeLa cells that harbors one to two rep-cap gene copies per cell. |
| Clinical Data |
31 Black female |
| Genes Expressed |
Rep proteins (Rep 78, Rep 68, Rep 52 and Rep 40), Cap proteins (VP1, VP2 and VP3) |
| Cellular Products |
Rep proteins (Rep 78, Rep 68, Rep 52 and Rep 40) Cap proteins (VP1, VP2 and VP3) |
| Comments |
This is a clone of HeLa cells that harbors one to two rep-cap gene copies per cell. Upon vector transfection and adenovirus infection, efficient rAAV assembly correlated with a 100-fold amplification of the integrated rep-cap sequence with the inverted terminal repeats (ITRs) deleted.
|
| 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%.
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| Subculturing |
Volumes used in this protocol are for 75 cm 2 flasks; 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 Ca++/Mg++ free Dulbecco's phosphate-buffered saline (D-PBS) or 0.25% (w/v) Trypsin - 0.53 mM EDTA solution to remove all traces of serum which contains trypsin inhibitor.
- Add 1.0 to 2.0 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.
- Transfer cell suspension to a centrifuge tube and spin at approximately 125 X g for 5 to 10 minutes. Discard supernatant.
- Resuspend the cell pellet in fresh growth medium. Add appropriate aliquots of the cell suspension to new culture vessels.
- Incubate cultures at 37°C.
Subcultivation ratio: A subcultivation ratio of 1:2 to 1:6 is recommended.
Medium renewal: Every 2 to 3 days |
| Cryopreservation |
Freeze medium: Dulbecco's Modified Eagle's Medium, 70%; fetal bovine serum, 20%; DMSO, 10% |
| STR Profile |
D5S818: 11, 12 D13S317: 12, 13.3 D7S820: 8, 12 D16S539: 9, 10 vWA: 16, 18 THO1: 7 TPOX: 8, 12 CSF1PO: 9, 10 Amelogenin: X |
| Name of Depositor |
P Moullier |
| References |
Chadeuf G, et al. Efficient recombinant adeno-associated virus production by a stable rep-cap HeLa cell line correlates with adenovirus-induced amplification of the integrated rep-cap genome. J. Gene Med. 2(4):260-268, 2000. PubMed: 10953917.
Tessier J, et al. Characterization of adenovirus-induced inverted terminal repeat-independent amplification of integrated adeno-associated virus rep-cap sequences. J. Virol. 75(1):375-383, 2001. PubMed: 11119606
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