transfection host (technology from amaxa
Roche FuGENE® Transfection Reagents)

negative

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: ].
Tested and found negative for ectromelia virus (mousepox).
This line is also designated as ATCC CCL-92.1.

Protocol: Never allow culture to become completely confluent.

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 that 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 37C 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.
   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.
6. Incubate cultures at 37C.

Interval: Every three days

Medium renewal: 2 to 3 times per week

14 hrs

886: Green H , Meuth M . An established pre-adipose cell line and its differentiation in culture. Cell 3: 127-133, 1974. PubMed:
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:
32455: Scherer PE , et al. Identification, sequence, and expression of caveolin-2 defines a caveolin gene family. Proc. Natl. Acad. Sci. USA 93: 131-135, 1996. PubMed:
32787: Kallen CB , Lazar MA . Antidiabetic thiazolidinediones inhibit leptin (ob) gene expression in 3T3-L1 adipocytes. Proc. Natl. Acad. Sci. USA 93: 5793-5796, 1996. PubMed: