| 產品名稱 |
GH354 |
| 商品貨號 |
B180442 |
| Organism |
Homo sapiens, human |
| Tissue |
cervix |
| Cell Type |
epithelial |
| Product Format |
frozen |
| Morphology |
epithelial |
| Culture Properties |
adherent |
| Biosafety Level |
2 [cells contain HPV-18 and Ad 5 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 |
adenocarcinoma |
| Age |
31 yrs adult |
| Gender |
female |
| Ethnicity |
Black |
| Applications |
This cell line can be used for the generation of E1-deleted adenovirus vectors. |
| Storage Conditions |
liquid nitrogen vapor phase |
| Derivation |
This line was derived from HeLa (ATCC CCL-2) [U.S. Pat. 6,365,394]. The line was stably transfected with plasmid constructs carrying a 3.4 kb DNA fragment of Ad 5 genome spanning 511 to 3924 bp (E1a and E1b open reading frames and part of the pIX gene) under the control of a phosphoglycerate kinase (PGK) promotor. The cells contain multiple copies of the plasmids, which carry multiple copies of E1a and E1b genes. The plasmid also contains the neomycin resistance gene.
The GH354 cell line was deposited at the ATCC as PTA-3404. Progeny from PTA-3404 are available as ATCC CRL-13003. |
| Clinical Data |
female Black 31 yrs adult |
| Comments |
The cells contain multiple copies of the plasmids, which carry multiple copies of E1a and E1b genes. The plasmid also contains the neomycin resistance gene.
The GH354 cell line was deposited at the ATCC as PTA-3404. Progeny from PTA-3404 are available as ATCC CRL-13003. |
| 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 20%.
|
| 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.
Subcultivation Ratio: 1:4 to 1:6
Medium Renewal: Every two to three 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% |
| Name of Depositor |
Trustees of Univ. of Pennsylvania |
| Deposited As |
Homo sapiens |
| References |
Gao GP, et al. A cell line for high-yield production of E1-deleted adenovirus vectors without the emergence of replication-competent virus. Hum. Gene Ther. 11: 213-219, 2000. PubMed: 10646652
Gao G, Wilson JM. Cell lines and constructs useful in production of E1-deleted adenoviruses in absence of replication competent adenovirus. US Patent 6,365,394 dated Apr 2 2002
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