GH354細(xì)胞

年限： 31 yrs *****

數(shù)量： 大量

細(xì)胞類型： 上皮細(xì)胞

運(yùn)輸方式： 凍存運(yùn)輸

是否是腫瘤細(xì)胞： 0

物種來(lái)源： 人

ATCC Number： CRL-13003?

相關(guān)**： 腺癌

細(xì)胞形態(tài)： 上皮樣

器官來(lái)源： 宮頸

生長(zhǎng)狀態(tài)： 貼壁生長(zhǎng)

GH354細(xì)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(xì)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(xì)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(xì)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