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Cell Biology
ATCC® Number: CRL-2972™    Price: $338.00
Designations: HeLaRC32 [HeRC32]
Depositors:  P Moullier
Biosafety Level: 2
Shipped: frozen
Medium & Serum: See Propagation
Growth Properties: adherent
Organism: Homo sapiens (human)
Morphology: epithelial
Source: Tissue: cervix
Disease: adenocarcinoma
Cellular Products: Rep proteins (Rep 78, Rep 68, Rep 52 and Rep 40)
Cap proteins (VP1, VP2 and VP3)
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. 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.
DNA Profile (STR): 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
Age: 31
Gender: female
Ethnicity: Black
Comments: 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. [PubMed: 11119606]. 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. These cells generate rAAV preparations with high titers of infectious particles which are essentially free of adenoviruses for biological, preclinical, clinical or pharmaceutical applications. [PubMed: 10953917].
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 10%.
Subculturing: Protocol: Volumes used in this protocol are for 75 sq cm 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 37C 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 37C.
    Subcultivation ratio: A subcultivation ratio of 1:2 to 1:6 is recommended.
    Medium renewal: Every 2 to 3 days
Preservation: Freeze medium: Dulbecco's Modified Eagle's Medium, 70%; fetal bovine serum, 20%; DMSO, 10%
Related Products: Recommended medium (without the additional serum described under ATCC Medium): ATCC 30-2002
Recommended serum: ATCC 30-2020
0.25% (w/v) Trypsin - 0.53mM EDTA in Hank's BSS (w/o Ca++, Mg ++): ATCC 30-2101
Phosphate-buffered saline: ATCC 30-2200
Cell culture tested DMSO: ATCC 4-X
Erythrosin B vital stain solution: ATCC 30-2404
Recommended adenovirus: ATCC VR-1616, rAAV2
Recommended vector: ATCC MBA-331, pTR-UF-
References: 16172640: 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.
16172684: 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
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