Growth

Properties:

adherent

Organism:

Mus musculus （mouse）

Source:

Organ: embryo
Cell type: fibroblast

Cellular Products:

triglycerides [3491]

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.

Propagation:

ATCC complete growth medium: RPMI-1640+10%FBS

Temperature: 37.0C
Atmosphere: air, 95%; carbon dioxide （CO2）, 5%
Growth Conditions: The cells should be grown in plastic flasks, they do not grow well on some types of glass surfaces. A saturation density of approximay 50000 cells per sq cm can be reached.

Subculturing:

Protocol: Never allow culture to become compley 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

Preservation:

Freeze medium: Complete growth medium supplemented with 5% （v/v） DMSO
Storage temperature: liquid nitrogen vapor phase

Doubling Time:

14 hrs

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
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: 8552590
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: 8650171