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Disease
tumor
Gender
male
Strain
Wistar
Storage Conditions
liquid nitrogen vapor phase
Derivation
The line was derived from a tumor which was induced with N-butyl-N-(4-hydroxybutyl) nitrosamine.
Comments
NBT-II cells differentiate in vitro giving rise to tumor giant cells.
The differentiation is inhibited by vitamin A.
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.
Remove and discard culture medium.
Briefly rinse the cell layer with 0.25% (w/v) Trypsin-0.03% (w/v) EDTA solution (ATCC 30-2101) 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 37°C 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 to new culture vessels.
Incubate cultures at 37°C.
Subcultivation Ratio: A subcultivation ratio of 1:4 to 1:10 is recommended Medium Renewal: 2 to 3 times per 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
Complete growth medium 95%; DMSO, 5%
Culture Conditions
Temperature: 37°C
Name of Depositor
J Leighton, R Tchao
Deposited As
Rattus sp.
References
Toyoshima K, et al. Tissue culture of urinary bladder tumor induced in a rat by N-butyl-N-(- 4-hydroxybutyl)nitrosamine: establishment of cell line, Nara Bladder Tumor II. J. Natl. Cancer Inst. 47: 979-985, 1971. PubMed: 4941696
Geisinger KR, et al. Appearance of giant cells as a morphological response of rat bladder carcinoma cell line (Nara Bladder Tumor No. 2) to a continuous thermal gradient in tissue culture. Cancer Res. 38: 1223-1230, 1978. PubMed: 639055
Toyoshima K, Leighton J. Vitamin A inhibition of keratinization in rat urinary bladder cancer cell line Nara Bladder Tumor No. 2 in meniscus gradient culture. Cancer Res. 35: 1873-1879, 1975. PubMed: 1093679
Tchao R. Novel forms of epithelial cell motility on collagen and on glass surfaces. Cell Motil. 2: 333-341, 1982. PubMed: 7151140
