GH354細胞
年限： 31 yrs *****
數量： 大量
細胞類型： 上皮細胞
運輸方式： 凍存運輸
是否是腫瘤細胞： 0
物種來源： 人
ATCC Number： CRL-13003?
相關**： 腺癌
細胞形態： 上皮樣
器官來源： 宮頸
生長狀態： 貼壁生長
GH354細胞Designations: GH354
Depositors: Trustees of Univ. of Pennsylvania
Biosafety Level: 2 [cells contain HPV-18 and Ad 5 viral DNA sequences ]
Shipped: frozen
Medium & Serum: See Propagation
Growth Properties: adherent
Organism: Homo sapiens
Morphology: epithelial
Source: Organ: cervix
Disease: adenocarcinoma
Cell Type: epithelial
Permits/Forms: In addition to the MTA mentioned above, other ATCC and/or regulatory permits may be required for the transfer of this ATCC material. GH354細胞Anyone purchasing ATCC material is ultimately responsible for obtaining the permits. Please click here for information regarding the specific requirements for shipment to your location.
Applications: generation of E1-deleted adenovirus vectors
Age: 31 yrs *****
Gender: female
Ethnicity: Black
Comments: 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 CRL-13003.
Propagation: ATCC 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%.
Atmosphere: air, 95%; carbon dioxide (CO2), 5%
Temperature: 37.0°C
Subculturing: GH354細胞Protocol:
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
Preservation: Freeze medium: Complete growth medium supplemented with 5% (v/v) DMSO
Storage temperature: liquid nitrogen vapor phase
Related Products: Recommended medium (without the additional supplements or serum described under ATCC Medium):ATCC 30-2002
recommended serum:ATCC 30-2020
parental cell line:ATCC PTA-3404
References: 65707: 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
70158: Gao G, Wilson JM. GH354細胞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
