BALB/3T3 clone A31 (ATCC® CCL-163)

Organism: Mus musculus, mouse  /  Cell Type: fibroblast  /  Tissue: embryo  / 

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Organism Mus musculus, mouse
Tissue embryo
Cell Type fibroblast
Product Format frozen
Morphology fibroblast
Culture Properties adherent
Biosafety Level 1
Age embryo; 14 to 17 days gestation
Strain BALB/c
Applications
This cell line is a suitable transfection host.
Storage Conditions liquid nitrogen vapor phase
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.
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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.
Tumorigenic No, the cells were not tumorigenic in immunosuppressed mice, but did form colonies in semisolid medium.
Effects
No, in immunosuppressed mice
Yes, in semisolid medium
Virus Susceptibility Herpes simplex virus
Vesicular stomatitis virus
Virus Resistance {265A7263-515F-42F9-89FD-9B140DF21BEC}
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).
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: bovine calf serum to a final concentration of 10%.
Subculturing
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
    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
    Growth Conditions: 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.
    Name of Depositor S Aaronson
    Year of Origin 1968
    References

    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

    Anderson MT, et al. Simultaneous fluorescence-activated cell sorter analysis of two distinct transcriptional elements within a single cell using engineered green fluorescent proteins. Proc. Natl. Acad. Sci. USA 93: 8508-8511, 1996. PubMed: 8710900

    Biological evaluation of medical devices. Part 5: Tests for in vitro cytotoxicity. Sydney, NSW, Australia:Standards Australia;Standards Australia AS ISO 10993.5-2002.

    Biological evaluation of medical devices--Part 5: Tests for in vitro cytotoxicity. Geneva (Switzerland):International Organization for Standardization/ANSI;ISO ISO 10993-5:1999.

    Notice: Necessary PermitsPermits

    These permits may be required for shipping this product:

    • Customers located in the state of Hawaii will need to contact the Hawaii Department of Agriculture to determine if an Import Permit is required. A copy of the permit or documentation that a permit is not required must be sent to ATCC in advance of shipment.
    Basic Documentation
    Other Documentation
    References

    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

    Anderson MT, et al. Simultaneous fluorescence-activated cell sorter analysis of two distinct transcriptional elements within a single cell using engineered green fluorescent proteins. Proc. Natl. Acad. Sci. USA 93: 8508-8511, 1996. PubMed: 8710900

    Biological evaluation of medical devices. Part 5: Tests for in vitro cytotoxicity. Sydney, NSW, Australia:Standards Australia;Standards Australia AS ISO 10993.5-2002.

    Biological evaluation of medical devices--Part 5: Tests for in vitro cytotoxicity. Geneva (Switzerland):International Organization for Standardization/ANSI;ISO ISO 10993-5:1999.