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HeLa S3


HeLa S3 cells are a clonal derivative of the parent HeLa cell line (ATCC CCL-2). 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 and is a suitable transfection host. The cells are positive for keratin by immunoperoxidase staining. HeLa cells have been reported to contain human papilloma virus 18 (HPV-18) sequences.
Product category
Human cells
Homo sapiens, human
Uterus; Cervix
3D cell culture
Product format
Storage conditions
Vapor phase of liquid nitrogen
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ATCC determines the biosafety level of a material based on our risk assessment as guided by the current edition of Biosafety in Microbiological and Biomedical Laboratories (BMBL), U.S. Department of Health and Human Services. It is your responsibility to understand the hazards associated with the material per your organization’s policies and procedures as well as any other applicable regulations as enforced by your local or national agencies.

Cells contain Human papillomavirus type 18 (HPV-18) sequences

ATCC highly recommends that appropriate personal protective equipment is always used when handling vials. For cultures that require storage in liquid nitrogen, it is important to note that some vials may leak when submersed in liquid nitrogen and will slowly fill with liquid nitrogen. Upon thawing, the conversion of the liquid nitrogen back to its gas phase may result in the vial exploding or blowing off its cap with dangerous force creating flying debris. Unless necessary, ATCC recommends that these cultures be stored in the vapor phase of liquid nitrogen rather than submersed in liquid nitrogen.

Required Products

These products are vital for the proper use of this item and have been confirmed as effective in supporting functionality. If you use alternative products, the quality and effectiveness of the item may be affected.

Detailed product information


Specific 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.


Growth properties
Passage history
A culture at approximately passage 400 was submitted to the American Type Culture Collection in February, 1972.
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.
31 years
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.
Virus susceptibility
Vesicular stomatitis, Glasgow (Indiana)
Vesicular stomatitis, Orsay (Indiana)
Encephalomyocarditis virus
Human adenovirus 5
Genes expressed
keratin positive by immunoperoxidase staining
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.

Handling information

Unpacking and storage instructions
  1. Check all containers for leakage or breakage.
  2. Remove the frozen cells from the dry ice packaging and immediately place the cells at a temperature below ­-130°C, preferably in liquid nitrogen vapor, until ready for use.
Complete 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%.
95% Air, 5% CO2
Handling procedure

To insure the highest level of viability, thaw the vial and initiate the culture as soon as possible upon receipt. If upon arrival, continued storage of the frozen culture is necessary, it should be stored in liquid nitrogen vapor phase and not at -70°C.  Storage at -70°C will result in loss of viability.

  1. Thaw the vial by gentle agitation in a 37°C water bath. To reduce the possibility of contamination, keep the O-ring and cap out of the water.  Thawing should be rapid (approximately 2 minutes).
  2. Remove the vial from the water bath as soon as the contents are thawed, and decontaminate by dipping in or spraying with 70% ethanol. All of the operations from this point on should be carried out under strict aseptic conditions.
  3. Transfer the vial contents to a centrifuge tube containing  9.0 mL complete culture medium. and spin at approximately 125 x g for 5 to 7 minutes.
  4. Resuspend cell pellet with the recommended complete medium (see the specific batch information for the culture recommended dilution ratio). and dispense into a 25 cm2 or a 75 cm2 culture flask.  It is important to avoid excessive alkalinity of the medium during recovery of the cells.  It is suggested that, prior to the addition of the vial contents, the culture vessel containing the complete growth medium be placed into the incubator for at least 15 minutes to allow the medium to reach its normal pH (7.0 to 7.6).
  5. Incubate the culture at 37°C in a suitable incubator.  A 5% CO2 in air atmosphere is recommended if using the medium described on this product sheet.
Subculturing procedure
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
Reagents for cryopreservation
Complete growth medium supplemented with 5% (v/v) DMSO (ATCC 4-X)

Quality control specifications

Mycoplasma contamination
Not detected
Virus testing
Human papillomavirus (HPV): Detected
STR profiling
Amelogenin: X
CSF1PO: 9,10
D13S317: 13.3
D16S539: 9,10
D5S818: 11,12
D7S820: 8,12
TH01: 7
TPOX: 8,12
vWA: 16,18
D3S1358: 15,18
D21S11: 27,28
D18S51: 16
Penta_E: 7,17
Penta_D: 8,15
D8S1179: 12,13
FGA: 18,21
D19S433: 13,14
D2S1338: 17


Deposited as
Homo sapiens
TT Puck
Year of origin
Cross references
GenBank Y09723 H.sapiens mRNA for Miz-1 protein.
GenBank Y08698 H.sapiens mRNA for RanBP3 (59 kDa).
GenBank Y08699 H.sapiens mRNA for RanBP3, splice variant.
GenBank U25182 Human antioxidant enzyme AOE37-2 mRNA, complete cds.
GenBank U33822 Human tax1-binding protein TXBP181 mRNA, complete cds.
GenBank U43430 Human epsilon isoform 14-3-3 protein mRNA, complete cds.
GenBank Y08697 H.sapiens mRNA for RanBP3 small splice variant (53 kDa).
GenBank L15328 Saccharomyces cerevisiae RNA helicase gene, complete cds.
GenBank U33821 Homo sapiens tax1-binding protein TXBP151 mRNA, complete cds.
GenBank NM_003443 Homo sapiens zinc finger protein 151 (pHZ-67) (ZNF151), mRNA.
GenBank AF034102 Homo sapiens NBMPR-insensitive nucleoside transporter ei (ENT2) mRNA, complete cds.
GenBank NM_006024 Homo sapiens Tax1 (human T-cell leukemia virus type I) binding protein 1 (TAX1BP1), mRNA.

Legal disclaimers

Intended use
This product is intended for laboratory research use only. It is not intended for any animal or human therapeutic use, any human or animal consumption, or any diagnostic use.

The product is provided 'AS IS' and the viability of ATCC® products is warranted for 30 days from the date of shipment, provided that the customer has stored and handled the product according to the information included on the product information sheet, website, and Certificate of Analysis. For living cultures, ATCC lists the media formulation and reagents that have been found to be effective for the product. While other unspecified media and reagents may also produce satisfactory results, a change in the ATCC and/or depositor-recommended protocols may affect the recovery, growth, and/or function of the product. If an alternative medium formulation or reagent is used, the ATCC warranty for viability is no longer valid.  Except as expressly set forth herein, no other warranties of any kind are provided, express or implied, including, but not limited to, any implied warranties of merchantability, fitness for a particular purpose, manufacture according to cGMP standards, typicality, safety, accuracy, and/or noninfringement.


This product is intended for laboratory research use only. It is not intended for any animal or human therapeutic use, any human or animal consumption, or any diagnostic use. Any proposed commercial use is prohibited without a license from ATCC.

While ATCC uses reasonable efforts to include accurate and up-to-date information on this product sheet, ATCC makes no warranties or representations as to its accuracy. Citations from scientific literature and patents are provided for informational purposes only. ATCC does not warrant that such information has been confirmed to be accurate or complete and the customer bears the sole responsibility of confirming the accuracy and completeness of any such information.

This product is sent on the condition that the customer is responsible for and assumes all risk and responsibility in connection with the receipt, handling, storage, disposal, and use of the ATCC product including without limitation taking all appropriate safety and handling precautions to minimize health or environmental risk. As a condition of receiving the material, the customer agrees that any activity undertaken with the ATCC product and any progeny or modifications will be conducted in compliance with all applicable laws, regulations, and guidelines. This product is provided 'AS IS' with no representations or warranties whatsoever except as expressly set forth herein and in no event shall ATCC, its parents, subsidiaries, directors, officers, agents, employees, assigns, successors, and affiliates be liable for indirect, special, incidental, or consequential damages of any kind in connection with or arising out of the customer's use of the product. While reasonable effort is made to ensure authenticity and reliability of materials on deposit, ATCC is not liable for damages arising from the misidentification or misrepresentation of such materials.

Please see the material transfer agreement (MTA) for further details regarding the use of this product. The MTA is available at

Permits & Restrictions

Import Permit for the State of Hawaii

If shipping to the U.S. state of Hawaii, you must provide either an import permit or documentation stating that an import permit is not required. We cannot ship this item until we receive this documentation. Contact the Hawaii Department of Agriculture (HDOA), Plant Industry Division, Plant Quarantine Branch to determine if an import permit is required.



Curated Citations

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.

View All Curated Citations for this Product

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