B16-OVA Cell Line

Cat No.: NTCC501219

B16-OVA is an OVA-transfected clone derived from the murine melanoma cell line B16. The tumor cell line was cultured in vitro

in CM in the presence of geniticin (2 mg/mL) and hygromicin B (60 μg/mL).

Culture Medium:

DMEM +10% FBS(Gibco)+ geniticin (2 mg/mL) and hygromicin B (60 μg/mL)

Subculture：

Volumes are given for a 25 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 0.25% (w/v) Trypsin 0.53mM 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. Incubate cultures at 37°C.

Subcultivation Ratio: A subcultivation ratio of 1:2 to 1:8 is recommended

Medium Renewal: 2-3days