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Applications: transfection host （Nucleofection technology from Lonza
Roche FuGENE® Transfection Reagents）
Virus Susceptibility: SV40 （lytic growth）; SV40 tsA209 at 40C; SV40 mutants with deletions in the early region
Comments: This is an African green monkey kidney fibroblast-like cell line suitable for transfection by vectors requiring expression of SV40 T antigen. This line contains T antigen, retains complete permissiveness for lytic growth of SV40, supports the replication of ts A209 virus at 40C, and supports the replication of pure populations of SV40 mutants with deletions in the early region.The line was derived from the CV-1 cell line （ATCC ® CCL-70?） by transformation with an origin defective mutant of SV40 which codes for wild type T antigen. The cells contain a single integrated copy of the complete early region of the SV40 genome.
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%.
Atmosphere: air, 95%; carbon dioxide （CO2）, 5%
Temperature: 37.0°C
Subculturing: Protocol:
Remove and discard medium.
Briefly rinse the cell layer with 0.25% （w/v） Trypsin - 0.53 mM EDTA solution to remove all traces of serum which 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 5 to 10 min）.
Note: To avoid clumping do not agitate the cells by hitting or shaking the flask while waiting for the cells to detatch. Cells that are difficult to detatch 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.
Incubate cultures at 37C.
Subc*tion Ratio: A subc*tion ratio of 1:4 to 1:8 is recommended
Medium Renewal: 2 to 3 times per week
Preservation: Freeze medium: Complete growth medium 95%; DMSO, 5%
Storage temperature: liquid nitrogen vapor temperature
Related Products: Recommended medium （without the additional supplements or serum described under ATCC Medium）:ATCC 30-2002
recommended serum:ATCC 30-2020
parental cell line:ATCC CCL-70
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
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