| 產(chǎn)品名稱 |
WEHI-13VAR |
| 商品貨號 |
B165939 |
| Organism |
Mus musculus, mouse |
| Product Format |
frozen |
| Morphology |
fibroblast |
| Culture Properties |
mixed, adherent and suspension |
| 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 |
fibrosarcoma |
| Strain |
BALB/c |
| Applications |
This line provides a stable and highly sensitive bioassay system to detect and measure mouse and human natural and recombinant tumor necrosis factors (TNF alpha and lymphotoxin. |
| Storage Conditions |
liquid nitrogen vapor phase |
| Derivation |
WEHI-13VAR, is a variant of WEHI 164 clone 13 which has lost its sensitivity to Tumor Necrosis Factor (TNF) in the absence of actinomycin D. |
| Comments |
The WEHI-13VAR cell line was found to maintain its high sensitivity to TNF when the assay was performed in the presence of 500 ng/mL actinomycin D. This cell line is more sensitive to TNF alpha and lymphotoxin than L929 (ATCC CCL-1) or WEHI 164 (ATCC CRL-1751). |
| Complete Growth Medium |
The base medium for this cell line is ATCC-formulated RPMI-1640 Medium, ATCC 30-2001. To make the complete growth medium, add the following components to the base medium: fetal bovine serum (ATCC 30-2020) to a final concentration of 10%.
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| 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-0.53 mM EDTA solution to remove all traces of serum that 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 37°C to facilitate dispersal.
- Add 6.0 to 8.0 mL of complete growth medium and aspirate cells by gently pipetting.
- To remove trypsin-EDTA solution, transfer cell suspension to the centrifuge tube with the medium and cells from Step 1 and spin at approximately 125 x g for 5 to 10 minutes.
- Discard supernatant and resuspend cells in fresh growth medium. Add appropriate aliquots of cell suspension to new culture vessels.
- Incubate cultures at 37°C.
Subculture Ratio: 1:4 to 1:6
Medium Renewal: Twice 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.
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| Cryopreservation |
Freeze medium: Complete growth medium supplemented with 5% (v/v) DMSO Storage temperature: liquid nitrogen vapor phase |
| Culture Conditions |
Temperature: 37.0°C |
| Name of Depositor |
JA Armstrong |
| Deposited As |
Mus musculus |
| References |
Espevik T, Nissen-Meyer J. A highly sensitive cell line, WEHI 164 clone 13, for measuring cytotoxic factor/tumor necrosis factor from human monocytes. J. Immunol. Methods 95: 99-105, 1986. PubMed: 3782828
Abu-Khabar KS, et al. Type I interferons (IFN-alpha and -beta) suppress cytotoxin (tumor necrosis factor-alpha and lymphotoxin) production by mitogen-stimulated human peripheral blood mononuclear cell. J. Leukocyte Biol. 52: 165-173, 1992. PubMed: 1506772
Khabar KS, et al. WEHI-13VAR: a stable and sensitive variant of WEHI 164 clone 13 fibrosarcoma for tumor necrosis factor bioassay. Immunol. Lett. 46: 107-110, 1995. PubMed: 7590904
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