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Cell Biology
ATCC® Number: CCL-2.2™    Price: $276.00
Designations: HeLa S3
Depositors:  TT Puck
Biosafety Level: 2 [Cells contain human papilloma virus (HPV-18) ]
Shipped: frozen
Medium & Serum: See Propagation
Growth Properties: adherent
Organism: Homo sapiens (human)
Morphology: epithelial
Source: Organ: cervix
Disease: adenocarcinoma
Cellular Products: keratin
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.
Isolation: Isolation date: 1955
Applications: transfection host (Roche FuGENE® Transfection Reagents
technology from amaxa)
DNA Profile (STR): Amelogenin: X
CSF1PO: 9,10
D13S317: 13.3
D16S539: 9,10
D5S818: 11,12
D7S820: 8,12
THO1: 7
TPOX: 8,12
vWA: 16,18
Cytogenetic Analysis: A medium-sized metacentric marker is present in 100% of the cells. HeLa Markers: One copy of M1, one copy of M2, two copies of M3, and one copy of M4.
Isoenzymes: G6PD, A
Age: 31 years
Gender: female
Ethnicity: Black
HeLa Markers: Y
Comments: HeLa S3 is a clonal derivative of the parent HeLa line (see ATCCCCL-2). S3 was cloned in 1955 by T.T. Puck, P.I. Marcus, and S.J. Cieciura. [22814]
The HeLa S3 clone has been very useful in the clonal analysis of mammalian cell populations relating to chromosomal variation, cell nutrition, and plaque-forming ability.
This line can be adapted to grow in suspension. [25952]
The cells are positive for keratin by immunoperoxidase staining.
A culture at approximately passage 400 was submitted to the American Type Culture Collection in February, 1972.
HeLa cells have been reported to contain human papilloma virus 18 (HPV-18) sequences. [23180]
Propagation: ATCC complete growth medium: The base medium for this cell line is ATCC-formulated F-12K Medium, Catalog No. 30-2004. To make the complete growth medium, add the following components to the base medium: fetal bovine serum to a final concentration of 10%.
Atmosphere: air, 95%; carbon dioxide (CO2), 5%
Temperature: 37.0°C
Subculturing: Protocol:
  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: A subcultivation ratio of 1:4 to 1:10 is recommended
Medium Renewal: 2 to 3 times per week
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 30-2004
recommended serum:ATCC 30-2020
References: 22263: Chen TR. Re-evaluation of HeLa, HeLa S3, and HEp-2 karyotypes. Cytogenet. Cell Genet. 48: 19-24, 1988. PubMed: 3180844
22766: Boshart M, et al. A new type of papillomavirus DNA, its presence in genital cancer biopsies and in cell lines derived from cervical cancer. EMBO J. 3: 1151-1157, 1984. PubMed: 6329740
22814: Puck TT, et al. Clonal growth of mammalian cells in vitro; growth characteristics of colonies from single HeLa cells with and without a feeder layer. J. Exp. Med. 103: 273-283, 1956. PubMed: 13286432
23180: Yee C, et al. Presence and expression of human papillomavirus sequences in human cervical carcinoma cell lines. Am. J. Pathol. 119: 361-366, 1985. PubMed: 2990217
25929: Puck TT, Marcus PI. A rapid method for viable cell titration and clone production with HeLa cells in tissue culture: the use of x-irradiated cells to supply conditioning factors. Proc. Natl. Acad. Sci. USA 41: 432-437, 1955.
25932: . . J. Exp. Med. 104: 427-434, 1956.
25934: . . Methods Enzymol. 5: 90-119, 1962.
25952: Darnell JE Jr., et al. The effect of cell population density on the amino acid requirements for poliovirus synthesis in HeLa cells. J. Exp. Med. 110: 445-450, 1959. PubMed: 13814142
25953: Cohen EP, Eagle H. A simplified chemostat for the growth of mammalian cells: characteristics of cell growth in continuous culture. J. Exp. Med. 113: 467-474, 1961.
25957: Darnell JE Jr., Sawyer TK. Variation in plaque-forming ability among parental and clonal strains of HeLa cells. Virology 8: 223-229, 1959. PubMed: 13669339
25959: Sato G, et al. Molecular growth requirements of single mammalian cells. Science 126: 461-464, 1957. PubMed: 13486039
32358: Soares K, et al. cis-Acting elements involved in transcriptional regulation of the herpes simples virus type 1 latency-associated promoter 1 (LAP1) in vitro and in vivo. J. Virol. 70: 5384-5394, 1996. PubMed: 8764049
32524: Chang YE, et al. Properties of the protein encoded by the UL32 open reading frame of herpes simplex virus 1. J. Virol. 70: 3938-3946, 1996. PubMed: 8648731
32966: Jiang BH, et al. Hypoxia-inducible factor 1 levels vary exponentially over a physiologically relevant range of O2 tension. Am. J. Physiol. 271: C1172-C1180, 1996. PubMed: 8897823
33006: Genuario RR, Perry RP. The GA-binding protein can serve as both an activator and repressor of ribosomal protein gene transcription. J. Biol. Chem. 271: 4388-4395, 1996. PubMed: 8626789
33032: Ladner RD, et al. Identification of a consensus cyclin-dependent kinase phosphorylation site unique to the nuclear form of human deoxyuridine triphosphate nucleotidohydrolase. J. Biol. Chem. 271: 7752-7757, 1996. PubMed: 8631817
33033: Ladner RD, et al. Characterization of distinct nuclear and mitochondrial forms of human deoxyuridine triphosphate nucleotidohydrolase. J. Biol. Chem. 271: 7745-7751, 1996. PubMed: 8631816
33034: Stewart L, et al. Biochemical and biophysical analyses of recombinant forms of human topoisomerase I. J. Biol. Chem. 271: 7593-7601, 1996. PubMed: 8631793
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