| Designations: | NIH/3T3 | | Biosafety Level: | 1 | | Shipped: | frozen | | Medium & Serum: | See Propagation | | Growth Properties: | adherent | | Organism: | Mus musculus （mouse） | | Morphology: | fibroblast
 | | Source: | Organ: embryo
Strain: NIH/Swiss
Cell Type: fibroblast fibroblast; | | Permits/Forms: | In addition to the MTA mentioned above, other ATCC and/or regulatory permits may be required for the transfer of this ATCC material. Anyone purchasing ATCC material is ultimay responsible for obtaining the permits. Please click here for information regarding the specific requirements for shipment to your location. |
| | Applications: | transfection host （Nucleofection technology from Lonza
Roche FuGENE? Transfection Reagents） | | Virus Susceptibility: | Murine leukemia virus | | Age: | embryo | | Comments: | The NIH/3T3 is highly sensitive to sarcoma virus focus formation and leukemia virus propagation and has proven to be very useful in DNA transfection studies [PubMed ID: 222457].
Tested and found negative for ectromelia virus （mousepox）. | | Propagation: | ATCC 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%.
Atmosphere: air, 95%; carbon dioxide （CO2）, 5%
Temperature: 37.0°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. | | Subculturing: | Protocol: - Remove and discard culture medium.
- 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.
- 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 37C to facilitate dispersal. - Add 6.0 to 8.0 ml of complete growth medium and aspirate cells by gently pipetting.
- Add appropriate aliquots of the cell suspension to new culture vessels.
- Incubate cultures at 37C.
.
DO NOT ALLOW THE CELLS TO BECOME CONFLUENT! Subculture at least twice per week at 80% confluence or less.
Subc*tion Ratio: Inoculate 3 to 5 X 10（3） cells/cm2
Medium Renewal: Twice per week | | Preservation: | Freeze medium: Complete growth medium supplemented with 5% （v/v） DMSO
Storage temperature: liquid nitrogen vapor phase | | Related Products: | Recommended medium （without the additional supplements or serum described under ATCC Medium）:ATCC 30-2002 | | References: | 22370: 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
26133: 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
26134: Copeland NG, Cooper GM. Transfection by exogenous and endogenous murine retrovirus DNAs. Cell 16: 347-356, 1979. PubMed: 222457
28301: Loffler S, et al. CD9, a tetraspan transmembrane protein, renders cells susceptible to canine distemper virus. J. Virol. 71: 42-49, 1997. PubMed: 8985321
32372: 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
32478: 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
32502: Gonzalez Armas JC, et al. DNA immunization confers protection against murine cytomegalovirus infection. J. Virol. 70: 7921-7928, 1996. PubMed: 8892915
32522: 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
32547: 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
32557: 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
32568: Lee JH, et al. The proximal promoter of the human transglutaminase 3 gene. J. Biol. Chem. 271: 4561-4568, 1996. PubMed: 8626812
32582: 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
32702: 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
32724: 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
32756: 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
32905: 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 |
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