Mu Islet (E6/E7) (ATCC® CRL-2999)

Organism: Mus musculus, mouse  /  Tissue: pancreas/islet cells; HPV16 E6/E7 transformed  / 

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Organism Mus musculus, mouse
Tissue pancreas/islet cells; HPV16 E6/E7 transformed
Product Format frozen
Morphology epithelial-like
Culture Properties adherent
Biosafety Level 2
Age 8 - 12 weeks
Gender male
Storage Conditions liquid nitrogen vapor phase
Derivation

CRL-2999 was established from isolated mouse Islets of Langerhans. The Islet cells were immortalized by transduction with pLXSN-E6E7 (ATCC CRL-2203) and selected with 400 µg/mL G418.

Comments
At ATCC, the cells have been shown to form small clusters when cultured in collagen gel. The cell clusters stain positive with Newport Green which would suggest some level of insulin expression.
Complete Growth Medium The base medium for this cell line is ATCC-formulated RPMI-1640 Medium, Catalog No. 30-2001. To make the complete growth medium, add the following components to the base medium: fetal bovine serum to a final concentration of 10%.
Subculturing
Volumes are given for a 75 cm2 flask. Increase or decrease the amount of dissociation medium needed proportionally for culture vessels of other sizes.
  1. Remove and discard culture medium.
  2. Briefly rinse the cell layer with Ca++/Mg++ free Dulbecco's phosphate-buffered saline (D-PBS) or 0.25% (w/v) Trypsin - 0.53 mM EDTA solution to remove all traces of serum which 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. An inoculum of 1 X 104 to 2 X 104 viable cells/cm2 is recommended.
  6. Incubate cultures at 37°C.

Subcultivation ratio: A subcultivation ratio of 1:3 to 1:4 is recommended.

Medium renewal: every 2 to 3 days

Cryopreservation
Freeze medium: complete growth medium, 95%; DMSO, 5%
Storage temperature: liquid nitrogen vapor phase
Culture Conditions
Temperature: 37°C
Atmosphere: air, 95%; carbon dioxide (CO2), 5%
Name of Depositor ATCC
Year of Origin March 2009
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