SNU-475 (ATCC® CRL-2236)

Organism: Homo sapiens, human  /  Tissue: liver  /  Disease: grade II-IV/V, hepatocellular carcinoma

Permits and Restrictions

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Organism Homo sapiens, human
Tissue liver
Product Format frozen
Morphology epithelial
Culture Properties adherent, monolayer of diffusely spreading cells
Biosafety Level 2  [Cells contain Hepatitis B virus]
Disease grade II-IV/V, hepatocellular carcinoma
Age 43 years
Gender male
Ethnicity Asian
Karyotype aneuploid; modal number = 61
Derivation
SNU-475 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.

Clinical Data
43 years
Asian
male

Antigen Expression
Blood Type AB; Rh +
Comments
Grossly, the original tumor was massive growth type. Histologically, it was predominantly compact and minor trabecular type.

The cultured cells are multinucleated.

Hepatitis B virus (HBV) DNA was detected by Southern blot hybridization.
HBV genomic RNA was not expressed.
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: heat-inactivated fetal bovine serum to a final concentration of 10%.
Subculturing Volumes used in this protocol are for 75 cm2 flask; 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 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

Subculture Ratio: 1:3 to 1:6
Medium Renewal: 2 to 3 timesa a week.  
Note: For more information on enzymatic dissociation and subculturing of cell lines consult Chapter 10 in Culture of Animal Cells, a manual of Basic Technique by R. Ian Freshney, 3rd edition, published by Alan R. Liss, N.Y., 1994.

Cryopreservation
Freeze medium: Complete growth medium, 95%; DMSO, 5%
Culture Conditions
Temperature: 37°C
STR Profile
Amelogenin: X,Y
CSF1PO: 11,12
D13S317: 8,11
D16S539: 12
D5S818: 10,13
D7S820: 7,12
THO1: 7,9
TPOX: 8,9
vWA: 14
Population Doubling Time 66 hrs
Name of Depositor J Park
Year of Origin 1990
References

Park JG, et al. Characterization of cell lines established from human hepatocellular carcinoma. Int. J. Cancer 62: 276-282, 1995. PubMed: 7543080

Basic Documentation
Other Documentation
Restrictions

This line is available under the following restrictions: 1.) The cell line was deposited for research purposes only. Neither the cell line nor products derived from it may be sold or used for commercial purposes. Nor can the cells be distributed to third parties for purpose of sale, or producing for sale, cells or their products. The cells are provided as a service to the research community. They are provided without warranty or merchantability of fitness for a particular purpose or any other warranty, express or implied. 2.) Any proposed commercial use of these cells or products produced by them must first be negotiated with Jae-GaHB-Park, Director, Korean Cell Line Bank, 28 Yongon-dong, Chongno-gu, Seoul, 110-744 Korea. Telephone (02) 760-3380, Fax (02) 742-4727. 3.) In all papers reporting any use of these cells or derived products, a direct reference will be made to the original publication (Int. J. Cancer 62:276-282, 1995).

References

Park JG, et al. Characterization of cell lines established from human hepatocellular carcinoma. Int. J. Cancer 62: 276-282, 1995. PubMed: 7543080