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Applications:
transfection host （Nucleofection technology from Lonza Roche FuGENE® Transfection Reagents）
Receptors:
insulin, expressed
Reverse Transcript:
negative
Age:
embryo
Comments:
L1 is a continuous substrain of 3T3 （Swiss albino） developed through clonal isolation. The cells undergo a pre-adipose to adipose like conversion as they progress from a rapidly dividing to a confluent and contact inhibited state. A high serum content in the medium enhances fat accumulation [PubMed ID: 4426090]. Tested and found negative for ectromelia virus （mousepox）. This line is also designated as ATCC CCL-92.1.ATCC CL-173 was deposited in 1974 without passage number information from the depositor. At the time of submission, ATCC prepared approximay 30 vials of seed stock at about 4 passages beyond the original depositor material （passage number: unknown +4）.
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.
Subculturing:
Protocol: Never allow culture to become compley confluent.
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 that 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. 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.
Incubate cultures at 37C.
Interval: Every three days Medium Renewal: 2 to 3 times per week
Preservation:
Freeze medium: Complete growth medium supplemented with 5% （v/v） DMSO Storage temperature: liquid nitrogen vapor phase
Doubling Time:
14 hrs
Related Products:
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
References:
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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