YS001 (ATCC® CRL-11776)

Organism: Mus musculus, mouse  /  Cell Type: embryonic stem cell  / 

Organism Mus musculus, mouse
Cell Type embryonic stem cell
Product Format frozen
Morphology epithelial
Culture Properties adherent
Biosafety Level 1
Age embryo, blastocyst embryo, blastocyst
Storage Conditions liquid nitrogen vapor phase
Complete Growth Medium Dulbecco's modified Eagle's medium with 4 mM L-glutamine adjusted to contain 1.5 g/L sodium bicarbonate and 4.5 g/L glucose and supplemented with 0.1 mM 2-mercaptoethanol, 85%; fetal bovine serum, 15%
  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.
  6. Incubate cultures at 37°C.
Subcultivation Ratio: A subcultivation ratio of 1:6 to 1:20 is recommended
Medium Renewal: Every 2 to 3 days
Freeze medium: Complete growth medium supplemented with 5% (v/v) DMSO
Storage temperature: liquid nitrogen vapor phase
Culture Conditions
Atmosphere: air, 95%; carbon dioxide (CO2), 5%
Temperature: 37.0°C
Growth Conditions: The line should be grown on feeder layers of irradiated (3000 rads) or mitomycin C treated (0.01 mg/ml for 90 minutes) primary mouse embryonic fibroblasts or STO cells (see ATCC CRL-1503 or ATCC 56-X, irradiated STO cells).
Name of Depositor TW Mak, Amgen, Inc.