Hs 695T (ATCC® HTB-137)

Organism: Homo sapiens, human  /  Cell Type: Melanoma  /  Tissue: skin; derived from metastatic site: lymph node  /  Disease: melanoma, amelanotic

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Organism Homo sapiens, human
Tissue skin; derived from metastatic site: lymph node
Cell Type Melanoma
Product Format frozen
Morphology epithelial
Culture Properties adherent
Biosafety Level 1
Disease melanoma, amelanotic
Age 26 years adult
Gender male
Ethnicity Caucasian
Storage Conditions liquid nitrogen vapor phase
Karyotype (P19-40) mode = 52; Y chromosome present
Derivation
Hs 695T was isolated in 1973 from the lymph node metastasis of an amelanotic melanoma by A. Creasey et al. 
Clinical Data male
Caucasian
26 years old
Tumorigenic Yes
Effects Yes, in immunosuppressed mice
Comments
The cells had a doubling time of approximately 48 hours at passages 19 through 40 and grew moderately in semi-solid medium.
Complete Growth Medium The base medium for this cell line is ATCC-formulated Eagle's Minimum Essential Medium, Catalog No. 30-2003. 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 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-0.53mM 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: 1:2 to 1:4
Medium Renewal: Every 2 to 3 days

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
Complete growth medium supplemented with 5% (v/v) DMSO. Cell culture tested DMSO is available as ATCC Catalog No. 4-X.

Culture Conditions

Temperature: 37°C
Atmosphere: 5% CO

STR Profile
Amelogenin: X,Y
CSF1PO: 11
D13S317: 12
D16S539: 9,13
D5S818: 9
D7S820: 9,10
THO1: 6
TPOX: 8
vWA: 18
Isoenzymes
AK-1, 1
ES-D, 1
G6PD, B
GLO-I, 1
Me-2, 0
PGM1, 1
PGM3, 1
Name of Depositor RB Owens
Passage History
Hs 695T was isolated in 1973 from the lymph node metastasis of an amelanotic melanoma by A. Creasey et al. The cells had a doubling time of approximately 48 hours at passages 19 through 40 and grew moderately in semi-solid medium.
Year of Origin 1973
References

Creasey AA, et al. Biological properties of human melanoma cells in culture. In Vitro 15: 342-350, 1979. PubMed: 478563

Hay, R. J., Caputo, J. L., and Macy, M. L., Eds. (1992), ATCC Quality Control Methods for Cell Lines. 2nd edition, Published by ATCC.

Caputo, J. L., Biosafety procedures in cell culture. J. Tissue Culture Methods 11:223-227, 1988.

Fleming, D.O., Richardson, J. H., Tulis, J.J. and Vesley, D., (1995) Laboratory Safety: Principles and Practice. Second edition, ASM press, Washington, DC.

Biosafety in Microbiological and Biomedical Laboratories, 5th ed. HHS. U.S. Department of Health and Human Services, Centers for Disease Control and Prevention. Washington DC: U.S. Government Printing Office; 2007. The entire text is available online at http://www.cdc.gov/OD/ohs/biosfty/bmbl5/bmbl5toc.htm

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
References

Creasey AA, et al. Biological properties of human melanoma cells in culture. In Vitro 15: 342-350, 1979. PubMed: 478563

Hay, R. J., Caputo, J. L., and Macy, M. L., Eds. (1992), ATCC Quality Control Methods for Cell Lines. 2nd edition, Published by ATCC.

Caputo, J. L., Biosafety procedures in cell culture. J. Tissue Culture Methods 11:223-227, 1988.

Fleming, D.O., Richardson, J. H., Tulis, J.J. and Vesley, D., (1995) Laboratory Safety: Principles and Practice. Second edition, ASM press, Washington, DC.

Biosafety in Microbiological and Biomedical Laboratories, 5th ed. HHS. U.S. Department of Health and Human Services, Centers for Disease Control and Prevention. Washington DC: U.S. Government Printing Office; 2007. The entire text is available online at http://www.cdc.gov/OD/ohs/biosfty/bmbl5/bmbl5toc.htm