GH354 (ATCC® CRL-13003)

Organism: Homo sapiens, human  /  Cell Type: epithelial  /  Tissue: cervix  /  Disease: adenocarcinoma

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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]
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.
  1. Remove and discard culture medium.
  2. 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.
  3. 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.
  4. Add 6.0 to 8.0 mL of complete growth medium and aspirate cells by gently pipetting.
  5. Add appropriate aliquots of the cell suspension to new culture vessels.
  6. 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
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

Notice: Necessary PermitsPermits

These permits may be required for shipping this product:

  • Customers located in the state of Hawaii will need to contact the Hawaii Department of Agriculture to determine if an Import Permit is required. A copy of the permit or documentation that a permit is not required must be sent to ATCC in advance of shipment.
Basic Documentation
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