NIH/3T3 (ATCC® CRL-1658)

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

Permits and Restrictions

View Permits

Organism Mus musculus, mouse
Tissue embryo
Cell Type fibroblast
Product Format frozen
Morphology fibroblast
Culture Properties adherent
Biosafety Level 1
Age embryo
Strain NIH/Swiss
Applications
This cell line is a suitable transfection host.
The NIH/3T3 cell line is highly sensitive to sarcoma virus focus formation and leukemia virus propagation and has proven to be very useful in DNA transfection studies. RefCopeland NG, Cooper GM. Transfection by exogenous and endogenous murine retrovirus DNAs. Cell 16: 347-356, 1979. PubMed: 222457 
Storage Conditions liquid nitrogen vapor phase
Images
Virus Susceptibility Murine leukemia virus , Murine leukemia virus
Comments
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.
  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 which 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.

DO NOT ALLOW THE CELLS TO BECOME CONFLUENT Subculture at least twice per week at 80% confluence or less.
Subcultivation Ratio: Inoculate 3 to 5 X 103 cells/cm2
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 important in culturing this line. Calf serum is recommended and not fetal bovine serum. The calf serum initially employed and found to be satisfactory was from the Colorado Serum Co. Denver.
References

Jainchill JL, et al. Murine sarcoma and leukemia viruses: assay using clonal lines of contact-inhibited mouse cells. J. Virol. 4: 549-553, 1969. PubMed: 4311790

Andersson P, et al. A defined subgenomic fragment of in vitro synthesized Moloney sarcoma virus DNA can induce cell transformation upon transfection. Cell 16: 63-75, 1979. PubMed: 84715

Copeland NG, Cooper GM. Transfection by exogenous and endogenous murine retrovirus DNAs. Cell 16: 347-356, 1979. PubMed: 222457

Loffler S, et al. CD9, a tetraspan transmembrane protein, renders cells susceptible to canine distemper virus. J. Virol. 71: 42-49, 1997. PubMed: 8985321

Berson JF, et al. A seven-transmembrane domain receptor involved in fusion and entry of T-cell-tropic human immunodeficiency virus tyep 1 strains. J. Virol. 70: 6288-6295, 1996. PubMed: 8709256

Jones PL, et al. Tumor necrosis factor alpha and interleukin-1beta regulate the murine manganese superoxide dismutase gene through a complex intronic enhancer involving C/EBP-beta and NF-kappaB. Mol. Cell. Biol. 17: 6970-6981, 1997. PubMed: 9372929

Gonzalez Armas JC, et al. DNA immunization confers protection against murine cytomegalovirus infection. J. Virol. 70: 7921-7928, 1996. PubMed: 8892915

Siess DC, et al. Exceptional fusogenicity of chinese hamster ovary cells with murine retrovirus suggests roles for cellular factor(s) and receptor clusters in the membrane fusion process. J. Virol. 70: 3432-439, 1996. PubMed: 8648675

Jang SI, et al. Activator protein 1 activity is involved in the regulation of the cell type-specific expression from the proximal promoter of the human profilaggrin gene. J. Biol. Chem. 271: 24105-24114, 1996. PubMed: 8798649

Medin JA, et al. Correction in trans for Fabry disease: expression, secretion, and uptake of alpha-galactosidase A in patient-derived cells driven by a high-titer recombinant retroviral vector. Proc. Natl. Acad. Sci. USA 93: 7917-7922, 1996. PubMed: 8755577

Lee JH, et al. The proximal promoter of the human transglutaminase 3 gene. J. Biol. Chem. 271: 4561-4568, 1996. PubMed: 8626812

Chang K, Pastan I. Molecular cloning of mesothelin, a differentiation antigen present on mesothelium, mesotheliomas, and ovarian cancers. Proc. Natl. Acad. Sci. USA 93: 136-140, 1996. PubMed: 8552591

Cranmer LD, et al. Identification, analysis, and evolutionary relationships of the putative murine cytomegalovirus homologs of the human cytomegalovirus UL82 (pp71) and UL83 (pp65) matrix phosphoproteins. J. Virol. 70: 7929-7939, 1996. PubMed: 8892916

Shisler J, et al. Induction of susceptibility to tumor necrosis factor by E1A is dependent on binding to either p300 or p105-Rb and induction of DNA synthesis. J. Virol. 70: 68-77, 1996. PubMed: 8523594

Cavanaugh VJ, et al. Murine cytomegalovirus with a deletion of genes spanning HindIII-J and -I displays altered cell and tissue tropism. J. Virol. 70: 1365-1374, 1996. PubMed: 8627652

Westerman KA, Leboulch P. Reversible immortalization of mammalian cells mediated by retroviral transfer and site-specific recombination. Proc. Natl. Acad. Sci. USA 93: 8971-8976, 1996. PubMed: 8799138

The NIH/3T3, a continuous cell line of highly contact-inhibited cells was established from NIH Swiss mouse embryo cultures in the same manner as the original random bred 3T3 (ATCC CCL-92) and the inbred BALB/c 3T3 (ATCC CCL-163). The established NIH/3T3 line was subjected to more than 5 serial cycles of subcloning in order to develop a subclone with morphologic characteristics best suited for transformation assays.

Cross References

Nucleotide (GenBank) : AF077527 Mus musculus syntenin mRNA, complete cds.

Nucleotide (GenBank) : AF104358 Mus musculus synectin mRNA, complete cds.

Nucleotide (GenBank) : NM_016807 Mus musculus syndecan binding protein (Sdcbp), mRNA.

Nucleotide (GenBank) : AF068116 Mus musculus eIF4E-like protein 4E-LP mRNA, complete cds.

Nucleotide (GenBank) : NM_019726 Mus musculus G protein pathway suppressor 2 (Gps2), mRNA.

Nucleotide (GenBank) : U83327 Mus musculus CC chemokine receptor-5 (CCR5) gene, complete cds.

Nucleotide (GenBank) : AF153696 Mus musculus G protein pathway suppressor 2 mRNA, complete cds.

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

Jainchill JL, et al. Murine sarcoma and leukemia viruses: assay using clonal lines of contact-inhibited mouse cells. J. Virol. 4: 549-553, 1969. PubMed: 4311790

Andersson P, et al. A defined subgenomic fragment of in vitro synthesized Moloney sarcoma virus DNA can induce cell transformation upon transfection. Cell 16: 63-75, 1979. PubMed: 84715

Copeland NG, Cooper GM. Transfection by exogenous and endogenous murine retrovirus DNAs. Cell 16: 347-356, 1979. PubMed: 222457

Loffler S, et al. CD9, a tetraspan transmembrane protein, renders cells susceptible to canine distemper virus. J. Virol. 71: 42-49, 1997. PubMed: 8985321

Berson JF, et al. A seven-transmembrane domain receptor involved in fusion and entry of T-cell-tropic human immunodeficiency virus tyep 1 strains. J. Virol. 70: 6288-6295, 1996. PubMed: 8709256

Jones PL, et al. Tumor necrosis factor alpha and interleukin-1beta regulate the murine manganese superoxide dismutase gene through a complex intronic enhancer involving C/EBP-beta and NF-kappaB. Mol. Cell. Biol. 17: 6970-6981, 1997. PubMed: 9372929

Gonzalez Armas JC, et al. DNA immunization confers protection against murine cytomegalovirus infection. J. Virol. 70: 7921-7928, 1996. PubMed: 8892915

Siess DC, et al. Exceptional fusogenicity of chinese hamster ovary cells with murine retrovirus suggests roles for cellular factor(s) and receptor clusters in the membrane fusion process. J. Virol. 70: 3432-439, 1996. PubMed: 8648675

Jang SI, et al. Activator protein 1 activity is involved in the regulation of the cell type-specific expression from the proximal promoter of the human profilaggrin gene. J. Biol. Chem. 271: 24105-24114, 1996. PubMed: 8798649

Medin JA, et al. Correction in trans for Fabry disease: expression, secretion, and uptake of alpha-galactosidase A in patient-derived cells driven by a high-titer recombinant retroviral vector. Proc. Natl. Acad. Sci. USA 93: 7917-7922, 1996. PubMed: 8755577

Lee JH, et al. The proximal promoter of the human transglutaminase 3 gene. J. Biol. Chem. 271: 4561-4568, 1996. PubMed: 8626812

Chang K, Pastan I. Molecular cloning of mesothelin, a differentiation antigen present on mesothelium, mesotheliomas, and ovarian cancers. Proc. Natl. Acad. Sci. USA 93: 136-140, 1996. PubMed: 8552591

Cranmer LD, et al. Identification, analysis, and evolutionary relationships of the putative murine cytomegalovirus homologs of the human cytomegalovirus UL82 (pp71) and UL83 (pp65) matrix phosphoproteins. J. Virol. 70: 7929-7939, 1996. PubMed: 8892916

Shisler J, et al. Induction of susceptibility to tumor necrosis factor by E1A is dependent on binding to either p300 or p105-Rb and induction of DNA synthesis. J. Virol. 70: 68-77, 1996. PubMed: 8523594

Cavanaugh VJ, et al. Murine cytomegalovirus with a deletion of genes spanning HindIII-J and -I displays altered cell and tissue tropism. J. Virol. 70: 1365-1374, 1996. PubMed: 8627652

Westerman KA, Leboulch P. Reversible immortalization of mammalian cells mediated by retroviral transfer and site-specific recombination. Proc. Natl. Acad. Sci. USA 93: 8971-8976, 1996. PubMed: 8799138

The NIH/3T3, a continuous cell line of highly contact-inhibited cells was established from NIH Swiss mouse embryo cultures in the same manner as the original random bred 3T3 (ATCC CCL-92) and the inbred BALB/c 3T3 (ATCC CCL-163). The established NIH/3T3 line was subjected to more than 5 serial cycles of subcloning in order to develop a subclone with morphologic characteristics best suited for transformation assays.