產品名稱 |
Clone M-3 [Cloudman S91 melanoma] |
商品貨號 |
B164274 |
Organism |
Mus musculus, mouse |
Tissue |
skin |
Cell Type |
melanocyte |
Product Format |
frozen |
Morphology |
epithelial |
Culture Properties |
adherent |
Biosafety Level |
1
Biosafety classification is based on U.S. Public Health Service Guidelines, it is the responsibility of the customer to ensure that their facilities comply with biosafety regulations for their own country. |
Disease |
melanoma |
Gender |
male |
Strain |
DBA |
Storage Conditions |
liquid nitrogen vapor phase |
Karyotype |
Stemline number is hypertetraploid. Karyotype is stable within stemline number. Marker chromosomes: A medium size chromosome with a submedian centromere and a smaller chromosome with a median centromere., The remaining 81 chromosomes have terminal centromeres, the first one is larger than normal. A minute chromosome was noted in 20% of the cells. |
Derivation |
Clone M-3, a melanin-producing cell line was adapted to cell culture by Y. Yasumura, A.H. Tashjian and G. Sato from a Cloudman S91 melanoma in a (C X DBA) F1 male mouse obtained from the Jackson Memorial Laboratory, Bar Harbor, Maine. |
Clinical Data |
male |
Genes Expressed |
melanin |
Cellular Products |
melanin |
Tumorigenic |
Yes |
Effects |
Yes, in C57BL mice (Melanotic tumors developed after inoculation of 10(4) cells in 5/6 mice.) |
Virus Susceptibility |
Herpes simplex virus
Vaccinia virus
Pseudorabies virus
Vesicular stomatitis, Glasgow (Indiana)
Vesicular stomatitis, Orsay (Indiana)
|
Virus Resistance |
poliovirus 1 |
Comments |
Tested and found negative for ectromelia virus (mousepox).
|
Complete Growth Medium |
The base medium for this cell line is ATCC-formulated F-12K Medium, Catalog No. 30-2004. To make the complete growth medium, add the following components to the base medium:
fetal bovine serum to a final concentration of 2.5%
horse serum to a final concentration of 15%
|
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.
- Remove and discard culture medium.
- Briefly rinse the cell layer with 0.25% (w/v) Trypsin 0.03% (w/v) EDTA solution to remove all traces of serum which contains trypsin inhibitor.
- 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 37oC to facilitate dispersal.
- Add 6.0 to 8.0 mL of complete growth medium and aspirate cells by gently pipetting.
- Add appropriate aliquots of the cell suspension into new culture vessels.
- Incubate cultures at 37°C.
Subcultivation Ratio: A subcultivation ratio of 1:3 to 1:4 is recommended
Medium Renewal: Every other day
|
Cryopreservation |
culture medium 95%; DMSO, 5% |
Culture Conditions |
Temperature: 37°C Atmosphere: air, 95%; carbon dioxide (CO2), 5% |
Name of Depositor |
G Sato |
Deposited As |
Mus musculus |
References |
Yasamura Y, et al. Establishment of four functional, clonal strains of animal cells in culture. Science 154: 1186-1189, 1966. PubMed: 4288399
Schmidt W, et al. Cell-free tumor antigen peptide-based cancer vaccines. Proc. Natl. Acad. Sci. USA 94: 3262-3267, 1997. PubMed: 9096381
|