NZP-29 (ATCC® CRL-1925)

Organism: Oryx dammah, oryx, short-horned  / 

Organism Oryx dammah, oryx, short-horned
Product Format frozen
Morphology epithelial
Culture Properties adherent
Biosafety Level 1
Storage Conditions liquid nitrogen vapor phase
Comments
The cells are pancytokeratin positive.
The cells can be propagated for at least 12 population doublings beyond frozen reference seed stock.
Complete Growth Medium Dulbecco's modified Eagle's medium, 45%; Ham's F12 medium, 45%; fetal bovine serum, 10%; plus the following additives: 10 ng/ml epidermal growth factor; 0.005 mg/ml insulin; 5 ng/ml selenium; 0.005 mg/ml transferrin
Subculturing
Volumes used in this protocol are for 75 cm2 flasks; proportionally reduce or increase amount of dissociation medium 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.
  6. Resuspend the cell pellet in fresh growth medium. Add appropriate aliquots of the cell suspension to new culture vessels.
  7. Incubate cultures at 37°C.
Subcultivation Ratio: A subcultivation ratio of 1:2 to 1:4 is recommended
Medium Renewal: Every 2 to 3 days
Cryopreservation 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°C
Name of Depositor L Munson