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GH354

規格:

價格:

編號:B180442

品牌:Mingzhoubio

標準菌株

定量菌液

DNA

RNA

規格:

凍干粉

斜面

甘油

平板

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產品名稱

GH354

商品貨號

B180442

Organism

Homo sapiens, human

Tissue

cervix

培養基

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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