Biosafety classification is based on U.S. Public Health Service Guidelines, it is the responsibility of the customer to ensure that their facilities comply with biosafety regulations for their own country.
Disease
normal
Age
adult
Gender
female
Applications
Viral vaccine production:
Virus propagation
Production of high-titer viral stocks
Rescue of clinical viral isolates
Detection of verotoxin
Efficacy testing
Malaria biology
Media testing
Mycoplasma testing
Substrate
Testing
Transfection host
Detection of virus in ground beef
Storage Conditions
liquid nitrogen vapor phase
Karyotype
This is a cell line with the hypodiploid chromosome count.
Comments
This STAT1 knockout Vero cell line was derived from the parental Vero cell line (ATCC® CCL-81™) at ATCC using CRISPR-Cas9 gene editing technology. This cell line carries a homozygous 199 nucleotide deletion spanning the third intron and the fourth exon of the STAT1 gene. This cell line does not express STAT1 protein.
STAT1 (Signal Transducer and Activator of Transcription 1) is a transcription factor required for the interferon-based cellular anti-viral response. The Vero.STAT1KO cell line (ATCC® CCL-81-VHG™) has been validated at the genomic, transcript, and protein bio-functional levels. It exhibits significant increased viral titer and enhanced virus production capability when compared to its parental cell line.
Complete Growth Medium
The base medium for this cell line is ATCC-formulated Eagle's Minimum Essential Medium (EMEM; ATCC 30-2003). To make the complete growth medium, add the following components to the base medium: Fetal Bovine Serum (FBS; ATCC 30-2020) to a final concentration of 10%.
Subculturing
Volumes used in this protocol are for 75 cm2 flask; proportionally reduce or increase amount of dissociation medium for culture vessels of other sizes. Corning® T-75 flasks (catalog #430641) are recommended for subculturing this product.
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 culture vessels.
Cultures can be established between 2 x 104 and 4 x 104 viable cells/cm2.
Incubate cultures at 37°C.
Interval: Maintain cultures at a cell concentration between 2 X 104 and 1.6 X 105 cell/cm2.
Subcultivation Ratio: A subcultivation ratio of 1:3 to 1:6 is recommended
Medium Renewal: 2 to 3 times per week
Cryopreservation
Complete growth medium supplemented with 5% (v/v) DMSO (ATCC 4-X)
Culture Conditions
Atmosphere: air, 95%; carbon dioxide (CO2), 5%
Temperature: 37°C
Cells per Vial
Approximately 2 x 106 cells
Volume
1.0 mL
Sterility Tests
Bacteria and yeast: No growth Mycoplasma: No growth
Functional Tests
Genotype testing for STAT1 knockout. Increased viral titer and enhanced viral production capability relative to parental Vero cell line (ATCC? CCL-81?).
Population Doubling Time
Approximately 28 hours
Name of Depositor
ATCC
Year of Origin
2019
References
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