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BALB/3T3 clone A31

CCL-163

BALB/3T3 clone A31 a cell line developed by S.A. Aaronson and G.T. Todaro in 1968 from disaggregated 14- to 17-day-old BALB/c embryos.
Product category
Animal cells
Organism
Mus musculus, mouse
Classification
Eukaryota, Animalia, Metazoa, Chordata, Vertebrata, Tetrapod
Cell type
fibroblast
Morphology
fibroblast
Tissue
Embryo
Applications
3D cell culture
Product format
Frozen
Storage conditions
Vapor phase of liquid nitrogen
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Documentation

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.

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

General

Specific applications
This cell line is a suitable transfection host.

Characteristics

Growth properties
Adherent
Derivation
The BALB/3T3 clone A31 is one of several cell lines (see ATCC CCL-164, BALB/3T12) developed by S.A. Aaronson and G.T. Todaro in 1968 from disaggregated 14- to 17-day-old BALB/c mouse embryos.
Age
14 to 17 days gestation
Strain
BALB/c
Karyotype
modal number = 78; range = 62 to 109. The stemline number is hypotetraploid. Most of the cells contained only telocentric or acrocentric chromosomes. Note: Cytogenetic information is based on initial seed stock at ATCC. Cytogenetic instability has been reported in the literature for some cell lines.
Tumorigenic
No;
No, in immunosuppressed mice
Yes, in semisolid medium
Virus susceptibility
Herpes simplex virus
Vesicular stomatitis virus
Comments
The cells are extremely sensitive to contact inhibition of cell division, grow at a high dilution, exhibit a low saturation density and are highly susceptible to transformation in tissue culture by the oncogenic DNA virus, SV40, and murine sarcoma virus
Tested and found negative for ectromelia virus (mousepox).

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 Dulbecco's Modified Eagle's Medium, Catalog No. 30-2002. To make the complete growth medium, add the following components to the base medium: bovine calf serum to a final concentration of 10%.
Temperature
37°C
Atmosphere
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. Discard supernatant.
  4. Resuspend the cell pellet with the recommended complete medium and dispense into a culture flask (see the specific batch information for the culture recommended dilution ratio).  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 are given for a 75 cm2 flask. Increase or decrease the amount of dissociation medium needed proportionally for culture vessels of other sizes. Corning T-75 flasks (catalog #430641) are recommended for subculturing this product.

Note: Never allow cultures to become completely confluent before subculture. 

Some important considerations in the handling of 3T3 cells: doubling time is about 18 hours in sparse cultures. The cells reach a saturation density of about 106 cells per 20 cm2. In order to maintain this property of high contact inhibition, it is necessary to transfer routinely at only high dilutions, otherwise variants tend to be selected having reduced contact inhibition. Such low density makes culture vessels appear sparse and cell growth sensitive to sub-optimal temperature and media conditions.

  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: For 60 mm plates, use an inoculum of 3 x 105 cells per plate and subculture every 3 days.
    Medium Renewal: Twice per week
    Note: The serum used is calf serum, NOT fetal calf serum. The depositor recommended calf serum because fetal calf serum causes transformation and loss of contact inhibition.

    Quality control specifications

    Mycoplasma contamination
    Not detected

    History

    Deposited as
    Mus musculus
    Depositors
    S Aaronson
    Year of origin
    1968

    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.
    Warranty

    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.

    Disclaimers

    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 www.atcc.org.

    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.

    MORE INFORMATION ABOUT PERMITS AND RESTRICTIONS

    Frequently Asked Questions

    References

    Curated Citations

    Aaronson SA, Todaro GJ. Development of 3T3-like lines from Balb-c mouse embryo cultures: transformation susceptibility to SV40. J. Cell. Physiol. 72: 141-148, 1968. PubMed: 4301006

    Todaro GJ, Aaronson SA. Properties of clonal lines of murine sarcoma virus transformed Balb-3T3 cells. Virology 38: 174-202, 1969. PubMed: 4306523

    Aaronson SA, Todaro GJ. Basis for the acquisition of malignant potential by mouse cells cultivated in vitro. Science 162: 1024-1026, 1968. PubMed: 4301647

    Jainchill JL, Todaro GJ. Stimulation of cell growth in vitro by serum with and without growth factor. Relation to contact inhibition and viral transformation. Exp. Cell Res. 59: 137-146, 1970. PubMed: 4194429

    Thompson SA, et al. COOH-terminal extended recombinant amphiregulin with bioactivity comparable with naturally derived growth factor. J. Biol. Chem. 271: 17927-17931, 1996. PubMed: 8663535

    View All Curated Citations for this Product

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