HT-144 (ATCC® HTB-63)

Organism: Homo sapiens, human  /  Tissue: malignant melanoma; derived from metastatic site: subcutaneous tissue  /  Disease: malignant melanoma

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Organism Homo sapiens, human
Tissue malignant melanoma; derived from metastatic site: subcutaneous tissue
Product Format frozen
Morphology fibroblast
Culture Properties adherent
Biosafety Level 1
Disease malignant melanoma
Age 29 years
Gender male
Ethnicity Caucasian
Storage Conditions liquid nitrogen vapor phase
Karyotype (P15) hypotriploid to hypertriploid (+A, +B, +C, +D, +E, +F, +G) with abnormalities including acrocentric fragments, minutes, secondary constrictions, breaks, large submetacentric, subtelocentric and minute markers
Derivation
This is one of an extensive series of human tumor lines isolated and characterized by J. Fogh.
Clinical Data
29 years
Caucasian
male
Antigen Expression
Blood Type O; Rh+; HLA A1, Aw24, B13, B15, Cw3, DRw4, DRw7
Tumorigenic Yes
Effects
Yes, in nude mice; forms anaplastic malignant tumor consistent with melanoma; tumors also form in steroid treated hamsters
Complete Growth Medium The base medium for this cell line is ATCC-formulated McCoy's 5a Medium Modified, Catalog No. 30-2007. 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- 0.53 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. Incubate cultures at 37°C.
Note: For more information on enzymatic dissociation and subculturing of cell lines consult Chapter 10 Culture of Animal Cells, a manual of Basic Technique by R. Ian Freshney, 3rd edition, published by Alan R. Liss, N.Y., 1994.  

Subcultivation Ratio: A subcultivation ratio of 1:3 to 1:8 is recommended
Medium Renewal: 2 to 3 times per week
Cryopreservation
Culture medium, 95%; DMSO, 5%
Culture Conditions
Atmosphere: air, 95%; carbon dioxide (CO2), 5%
Temperature: 37°C
STR Profile
Amelogenin: X,Y
CSF1PO: 12
D13S317: 11,12
D16S539: 12,13
D5S818: 11,13
D7S820: 11
THO1: 6,9
TPOX: 8,11
vWA: 16,18
Isoenzymes
AK-1, 1
ES-D, 1-2
G6PD, B
GLO-I, 2
PGM1, 1-2
PGM3, 1
Name of Depositor J Fogh
References

Fogh J. Human tumor cells in vitro. New York: Plenum Press; 1975.

Fogh J, et al. Absence of HeLa cell contamination in 169 cell lines derived from human tumors. J. Natl. Cancer Inst. 58: 209-214, 1977. PubMed: 833871

Goodfellow M, et al. One hundred and twenty-seven cultured human tumor cell lines producing tumors in nude mice. J. Natl. Cancer Inst. 59: 221-226, 1977. PubMed: 77210034

Pollack MS, et al. HLA-A, B, C and DR alloantigen expression on forty-six cultured human tumor cell lines. J. Natl. Cancer Inst. 66: 1003-1012, 1981. PubMed: 7017212

Wang R, et al. Cellular adherence elicits ligand-independent activation of the Met cell-surface receptor. Proc. Natl. Acad. Sci. USA 93: 8425-8430, 1996. PubMed: 8710887

Basic Documentation
Restrictions

The cells are distributed for research purposes only. The Memorial Sloan-Kettering Cancer Center releases the cells subject to the following: 1.) The cells or their products must not be distributed to third parties. Commercial interests are the exclusive property of Memorial Sloan-Kettering Cancer Center. 2.) Any proposed commercial use of these cells must first be negotiated with the Office of Technology Development, Memorial Sloan-Kettering Cancer Center, 1275 York Avenue, New York, NY 10065. Contact Tingting Zhang-Kharas, Direct Phone: 646-888-1083, Reception: 646-888-1080, Email: zhangkht@mskcc.org

References

Fogh J. Human tumor cells in vitro. New York: Plenum Press; 1975.

Fogh J, et al. Absence of HeLa cell contamination in 169 cell lines derived from human tumors. J. Natl. Cancer Inst. 58: 209-214, 1977. PubMed: 833871

Goodfellow M, et al. One hundred and twenty-seven cultured human tumor cell lines producing tumors in nude mice. J. Natl. Cancer Inst. 59: 221-226, 1977. PubMed: 77210034

Pollack MS, et al. HLA-A, B, C and DR alloantigen expression on forty-six cultured human tumor cell lines. J. Natl. Cancer Inst. 66: 1003-1012, 1981. PubMed: 7017212

Wang R, et al. Cellular adherence elicits ligand-independent activation of the Met cell-surface receptor. Proc. Natl. Acad. Sci. USA 93: 8425-8430, 1996. PubMed: 8710887