liver

SNU-449 was derived in 1990 by J.-G. Park and associates from a primary hepatocellular carcinoma taken from a Korean patient prior to cytotoxic therapy.

Tumor cells were initially cultured in ACL-4 medium supplemented with 5% heat inactivated fetal bovine serum. After establishment, cultures were maintained in RPMI 1640 supplemented with 10% heat-inactivated fetal bovine serum.

52 years

Asian

male

CRL-2234 SNU-449 人肝癌細(xì)胞

The cultured cells contain a single or double nucleus.

Protocol:

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 1:5 to 1:10 is recommended

Medium Renewal: Every 2 to 3 days    CRL-2234 SNU-449 人肝癌細(xì)胞

Freeze medium: Complete growth medium supplemented with 5% (v/v) DMSO

Storage temperature: liquid nitrogen vapor phase

Atmosphere: air, 95%; carbon dioxide (CO₂), 5%

Temperature: 37°C