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3T3-L1
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Massachusetts Institute of Technology
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Biosafety Level:
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1
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frozen
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Medium & Serum:
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See Propagation
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adherent
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Mus musculus (mouse)
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fibroblast
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Organ: embryo
Cell Type: fibroblast
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triglycerides [3491]
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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 ultimately responsible for obtaining the permits. Please click here for information regarding the specific requirements for shipment to your location.
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transfection host (Nucleofection technology from Lonza
Roche FuGENE® Transfection Reagents)
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insulin, expressed
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negative
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embryo
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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.
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Protocol: Never allow culture to become completely confluent.
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Remove and discard culture medium.
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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.
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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.
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Add 6.0 to 8.0 ml of complete growth medium and aspirate cells by gently pipetting.
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Add appropriate aliquots of the cell suspension to new culture vessels.
The recommended inoculum is 2 to 3 X 10(3) cells/sq. cm. Subculture before cultures become 70 to 80% confluent or when cells reach 5 to 6 X10(4) viable cells/sq. cm.
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Incubate cultures at 37C.
Interval: Every three days
Medium Renewal: 2 to 3 times per week
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Freeze medium: Complete growth medium supplemented with 5% (v/v) DMSO
Storage temperature: liquid nitrogen vapor phase
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14 hrs
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Recommended medium (without the additional supplements or serum described under ATCC Medium):ATCC 30-2002
formerly distributed as:ATCC CCL-92.1
0.25% (w/v) Trypsin - 0.53 mM EDTA in Hank' BSS (w/o Ca++, Mg++):ATCC 30-2101
Cell culture tested DMSO:ATCC 4-X
Recommended serum: ATCC 30-2030
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886: Green H, Meuth M. An established pre-adipose cell line and its differentiation in culture. Cell 3: 127-133, 1974. PubMed: 4426090
3491: Green H. Triglyceride-accumulating clonal cell line. US Patent 4,003,789 dated Jan 18 1977
32373: Goodrum FD, et al. Adenovirus early region 4 34-kilodalton protein directs the nuclear localization of the early region 1B 55-kilodalton protein in primate cells. J. Virol. 70: 6323-6335, 1996. PubMed: 8709260
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