| NIH/3T3 | | Biosafety Level: | 1 | | frozen | | See Propagation | | adherent | | Mus musculus （mouse） | | fibroblast
| | Organ: embryo
Strain: NIH/Swiss
Cell Type: fibroblast fibroblast; | | 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. | | | | transfection host （Nucleofection technology from Lonza
Roche FuGENE® Transfection Reagents） | | Murine leukemia virus | | embryo | | 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. | | 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 | | Freeze medium: Complete growth medium supplemented with 5% （v/v） DMSO
Storage temperature: liquid nitrogen vapor phase | | Recommended medium （without the additional supplements or serum described under ATCC Medium）:ATCC 30-2002 | | 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
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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
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