Cellular Products: plasminogen activator; laminin; type IV collagen
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Comments: F9 cells can be stimulated to differentiate into parietal endoderm in the presence of retinoic acid and dibutyryl cyclic AMP （cAMP）.
Differentiating cells synthesize plasminogen activator, laminin and type IV collagen.
cAMP is active only on cells that have been treated with retinoic acid.
The cells maintain three copies of the beta 1 integrin gene.
Tested and found negative for ectromelia virus （mousepox）.
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: fetal bovine serum to a final concentration of 10%.
Temperature: 37.0°C
Subculturing: Protocol: NOTE: culture vessels must be coated with 0.1% gelatin prior to use.To do so, cover the surface of the vessel with 0.1% gelatin （Difco） in sterile distilled water for 2 hours at 4C, then wash three times with sterile distilled water. Treated flasks and dishes can be stored at room temperature.
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 37°C 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 coated culture vessels.
Incubate cultures at 37°C.
Subc*tion Ratio: A subc*tion ratio of 1:10 is recommended
Medium Renewal: Every 2 to 3 days
Related Products: Recommended medium （without the additional supplements or serum described under ATCC Medium）:ATCC 30-2002
recommended serum:ATCC 30-2020
References: 1160: Strickland S, et al. Hormonal induction of differentiation in teratocarcinoma stem cells: generation of parietal endoderm by retinoic acid and dibutyryl cAMP. Cell 21: 347-355, 1980. PubMed: 6250719
1161: Strickland S, Mahdavi V. The induction of differentiation in teratocarcinoma stem cells by retinoic acid. Cell 15: 393-403, 1978. PubMed: 214238
23426: Stephens LE, et al. Targeted deletion of beta 1 integrins in F9 embryonal carcinoma cells affects morphological differentiation but not tissue-specific gene expression. J. Cell Biol. 123: 1607-1620, 1993. PubMed: 7504677
26151: Berstine EG, et al. Alkaline phosphatase activity in mouse teratoma. Proc. Natl. Acad. Sci. USA 70: 3899-3903, 1973. PubMed: 4521215
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
