13F3 (ATCC® CRL-3422)

Organism: Cricetulus migratorius, hamster, Armenian  /  Tissue: spleen  / 

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Organism Cricetulus migratorius, hamster, Armenian
Tissue spleen
Product Format frozen 1.0 mL
Morphology rounded
Culture Properties mixed
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.

Applications Model for a novel and structurally distinct Ig superfamily inhibitory ligand (VISTA), designated as V-domain Ig suppressor of T cell activation, a immunoregulatory molecule, that may play a role in the development of autoimmunity and immune surveillance in cancer.
Images CRL-3422
Comments Armenian hamsters were immunized with EL4 cells overexpressing VISTA-RFP and VISTA-Ig and fused to the myeloma cell line SP2/0-Ag14.
Complete Growth Medium The base medium for this cell line is RPMI-1640 (ATCC 30-2001). To make the complete medium add 10% Fetal Bovine Serum (FBS; ATCC 30-2020) and an additional 2mM L-Glutamine (ATCC 30-2214).
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 2.0 x 104 and 6.0 x 104 viable cells/cm2.
  6. Incubate cultures at 37°C.
Interval: Maintain cultures at a cell concentration between 2.0 X 104 and 6.4 X 105 cell/cm2.
Subcultivation Ratio: A subcultivation ratio of 1:3 to 1:8 is recommended
Medium Renewal: 2 to 3 times per week
Cryopreservation Culture medium + 5% DMSO (ATCC 4-X)
Culture Conditions
Atmosphere: air, 95%; carbon dioxide (CO2), 5%
Temperature: 37°C
Cells per Vial Approximately 5.0 to 7.0 x 106 cells
Volume 1.0 mL
Sterility Tests Bacteria and yeast: No growth
Mycoplasma: No growth
Viral Testing Hepatitis B: None detected
Cytomegalovirus: None detected
Human immunodeficiency virus: None detected
Epstein-Barr virus: None detected
Human papillomavirus: None detected
Name of Depositor Dartmouth College
Year of Origin 2011
References

Wang L, et al. VISTA, a novel mouse Ig superfamily ligand that negatively regulates T cell responses. J Exp Med 208(3):577-592, 2011. PubMed: 21383057

Notice: Necessary PermitsPermits

These permits may be required for shipping this product:

  • Customers located in the state of Hawaii will need to contact the Hawaii Department of Agriculture to determine if an Import Permit is required. A copy of the permit or documentation that a permit is not required must be sent to ATCC in advance of shipment.
Basic Documentation
Other Documentation
References

Wang L, et al. VISTA, a novel mouse Ig superfamily ligand that negatively regulates T cell responses. J Exp Med 208(3):577-592, 2011. PubMed: 21383057