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Vero.STAT1 KO
Vero.STAT1 KO
規(guī)格:
貨期:
編號(hào):B161701
品牌:Mingzhoubio

標(biāo)準(zhǔn)菌株
定量菌液
DNA
RNA

規(guī)格:
凍干粉
斜面
甘油
平板


產(chǎn)品名稱 Vero.STAT1 KO
商品貨號(hào) B161701
Organism Cercopithecus aethiops
Tissue kidney
Product Format frozen 1.0 mL
Morphology epithelial
Culture Properties adherent
Biosafety Level 1

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.
  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 37°C 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.
    Cultures can be established between 2 x 104 and 4 x 104 viable cells/cm2.
  6. 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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Meraz MA, et al. Targeted disruption of the Stat1 gene in mice reveals unexpected physiologic specificity in the JAK-STAT signaling pathway. Cell 84: 431-442, 1996. Pubmed: 860859

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