HeLaRC32 [HeRC32] (ATCC® CRL-2972)

Organism: Homo sapiens, human  /  Tissue: cervix  /  Disease: adenocarcinoma

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Organism Homo sapiens, human
Tissue cervix
Product Format frozen
Morphology epithelial
Culture Properties adherent
Biosafety Level 2  [Cells contain human papilloma virus 18 (HPV-18)]
Disease adenocarcinoma
Age 31
Gender female
Ethnicity Black
Applications

These cells generate rAAV preparations with high titers of infectious particles which are essentially free of adenoviruses for biological, preclinical, clinical or pharmaceutical applications.

HelaRC32 is a stable packaging cell line expressing the rep and cap genes for recombinant adeno-associated virus type 2 (rAAV-2) assembly which constitutes an attractive alternative to transient transfection protocols. RefTessier J, et al. Characterization of adenovirus-induced inverted terminal repeat-independent amplification of integrated adeno-associated virus rep-cap sequences. J. Virol. 75(1):375-383, 2001. PubMed: 11119606

Storage Conditions liquid nitrogen vapor phase
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Derivation
This is a clone of HeLa cells that harbors one to two rep-cap gene copies per cell.
Clinical Data
31
Black
female
Genes Expressed
Rep proteins (Rep 78, Rep 68, Rep 52 and Rep 40), Cap proteins (VP1, VP2 and VP3)
Cellular Products
Rep proteins (Rep 78, Rep 68, Rep 52 and Rep 40)
Cap proteins (VP1, VP2 and VP3)
Comments

This is a clone of HeLa cells that harbors one to two rep-cap gene copies per cell. Upon vector transfection and adenovirus infection, efficient rAAV assembly correlated with a 100-fold amplification of the integrated rep-cap sequence with the inverted terminal repeats (ITRs) deleted.

 

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 10%.
Subculturing
Volumes used in this protocol are for 75 cm2 flasks; proportionally reduce or increase amount of dissociation medium for culture vessels of other sizes.
  1. Remove and discard culture medium.
  2. Briefly rinse the cell layer with Ca++/Mg++ free Dulbecco's phosphate-buffered saline (D-PBS) or 0.25% (w/v) Trypsin - 0.53 mM EDTA solution to remove all traces of serum which contains trypsin inhibitor.
  3. Add 1.0 to 2.0 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. Transfer cell suspension to a centrifuge tube and spin at approximately 125 X g for 5 to 10 minutes. Discard supernatant.
  6. Resuspend the cell pellet in fresh growth medium. Add appropriate aliquots of the cell suspension to new culture vessels.
  7. Incubate cultures at 37°C.
Subcultivation ratio: A subcultivation ratio of 1:2 to 1:6 is recommended.
Medium renewal: Every 2 to 3 days
Cryopreservation
Freeze medium: Dulbecco's Modified Eagle's Medium, 70%; fetal bovine serum, 20%; DMSO, 10%
STR Profile
D5S818: 11, 12
D13S317: 12, 13.3
D7S820: 8, 12
D16S539: 9, 10
vWA: 16, 18
THO1: 7
TPOX: 8, 12
CSF1PO: 9, 10
Amelogenin: X
Name of Depositor P Moullier
References

Chadeuf G, et al. Efficient recombinant adeno-associated virus production by a stable rep-cap HeLa cell line correlates with adenovirus-induced amplification of the integrated rep-cap genome. J. Gene Med. 2(4):260-268, 2000. PubMed: 10953917.

Tessier J, et al. Characterization of adenovirus-induced inverted terminal repeat-independent amplification of integrated adeno-associated virus rep-cap sequences. J. Virol. 75(1):375-383, 2001. PubMed: 11119606

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
Other Documentation
References

Chadeuf G, et al. Efficient recombinant adeno-associated virus production by a stable rep-cap HeLa cell line correlates with adenovirus-induced amplification of the integrated rep-cap genome. J. Gene Med. 2(4):260-268, 2000. PubMed: 10953917.

Tessier J, et al. Characterization of adenovirus-induced inverted terminal repeat-independent amplification of integrated adeno-associated virus rep-cap sequences. J. Virol. 75(1):375-383, 2001. PubMed: 11119606