HeLa S3 (ATCC® CCL-2.2)

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

Organism Homo sapiens, human
Tissue cervix
Product Format frozen
Morphology epithelial
Culture Properties adherent
Biosafety Level 2 [Cells contain human papilloma virus (HPV-18)]
Disease adenocarcinoma
Age 31 years
Gender female
Ethnicity Black
Applications
This cell line is a suitable transfection host.

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.
Storage Conditions liquid nitrogen vapor phase
Karyotype 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. Note: Cytogenetic information is based on initial seed stock at ATCC. Cytogenetic instability has been reported in the literature for some cell lines.
Derivation
HeLa S3 is a clonal derivative of the parent HeLa line (see ATCC CCL-2). S3 was cloned in 1955 by T.T. Puck, P.I. Marcus, and S.J. Cieciura.
Clinical Data
31 years
Black
female
HeLa Markers Y
Genes Expressed
The cells are positive for keratin by immunoperoxidase staining.
keratin.
Cellular Products
keratin
Virus Susceptibility Vesicular stomatitis, Glasgow (Indiana)
Vesicular stomatitis, Orsay (Indiana)
Encephalomyocarditis virus
Human adenovirus 5
Virus Resistance Human poliovirus 1
Human poliovirus 2
Human poliovirus 3
Comments
This line can be adapted to grow in suspension.

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. ATCC confirmed this cell line is positive for the presence of Papillomavirus viral DNA sequences via PCR.

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%.
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. Flasks do not become 100% confluent. Cells are rounded and have a tendency to float in the medium.
  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
Cryopreservation
Freeze medium: Complete growth medium supplemented with 5% (v/v) DMSO
Storage temperature: liquid nitrogen vapor phase
Culture Conditions
Atmosphere: air, 95%; carbon dioxide (CO2), 5%
Temperature: 37°C
STR Profile
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
Isoenzymes
G6PD, A
Name of Depositor TT Puck
Passage History
A culture at approximately passage 400 was submitted to the American Type Culture Collection in February, 1972.
Year of Origin 1955
References

Chen TR. Re-evaluation of HeLa, HeLa S3, and HEp-2 karyotypes. Cytogenet. Cell Genet. 48: 19-24, 1988. PubMed: 3180844

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

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

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

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.

Puck TT, Fisher HW. Genetics of Somatic Mammalian Cells. J. Exp. Med. 104: 427-434, 1956. PubMed: 19867118

Ham RG, Puck TT. Quantitative colonial growth of isolated mammalian cells. Methods Enzymol 5 90-119 1962

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

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.

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

Sato G, et al. Molecular growth requirements of single mammalian cells. Science 126: 461-464, 1957. PubMed: 13486039

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

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

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

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

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

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

Stewart L, et al. Biochemical and biophysical analyses of recombinant forms of human topoisomerase I. J. Biol. Chem. 271: 7593-7601, 1996. PubMed: 8631793

Cross References

Nucleotide (GenBank) : Y09723 H.sapiens mRNA for Miz-1 protein.

Nucleotide (GenBank) : Y08698 H.sapiens mRNA for RanBP3 (59 kDa).

Nucleotide (GenBank) : Y08699 H.sapiens mRNA for RanBP3, splice variant.

Nucleotide (GenBank) : U25182 Human antioxidant enzyme AOE37-2 mRNA, complete cds.

Nucleotide (GenBank) : U33822 Human tax1-binding protein TXBP181 mRNA, complete cds.

Nucleotide (GenBank) : U43430 Human epsilon isoform 14-3-3 protein mRNA, complete cds.

Nucleotide (GenBank) : Y08697 H.sapiens mRNA for RanBP3 small splice variant (53 kDa).

Nucleotide (GenBank) : L15328 Saccharomyces cerevisiae RNA helicase gene, complete cds.

Nucleotide (GenBank) : U33821 Homo sapiens tax1-binding protein TXBP151 mRNA, complete cds.

Nucleotide (GenBank) : NM_003443 Homo sapiens zinc finger protein 151 (pHZ-67) (ZNF151), mRNA.

Nucleotide (GenBank) : AF034102 Homo sapiens NBMPR-insensitive nucleoside transporter ei (ENT2) mRNA, complete cds.

Nucleotide (GenBank) : NM_006024 Homo sapiens Tax1 (human T-cell leukemia virus type I) binding protein 1 (TAX1BP1), mRNA.

Basic Documentation
References

Chen TR. Re-evaluation of HeLa, HeLa S3, and HEp-2 karyotypes. Cytogenet. Cell Genet. 48: 19-24, 1988. PubMed: 3180844

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

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

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

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.

Puck TT, Fisher HW. Genetics of Somatic Mammalian Cells. J. Exp. Med. 104: 427-434, 1956. PubMed: 19867118

Ham RG, Puck TT. Quantitative colonial growth of isolated mammalian cells. Methods Enzymol 5 90-119 1962

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

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.

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

Sato G, et al. Molecular growth requirements of single mammalian cells. Science 126: 461-464, 1957. PubMed: 13486039

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

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

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

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

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

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

Stewart L, et al. Biochemical and biophysical analyses of recombinant forms of human topoisomerase I. J. Biol. Chem. 271: 7593-7601, 1996. PubMed: 8631793