| Applications |
AT3B-1 was derived from the AT-3 cell line. They are more resistant to vinblastine compared to controls. These cells express a multidrug (MDR) resistant phenotype. Injection of AT3 B-1 cells into rats followed by doxorubicin treatment produced larger tumors compared to the parental controls. When P-glycoprotein was blocked, the AT3 B-1 cell line demonstrated drug efflux pump activity. This cell line can be used to study chemotherapy resistance in prostate cancer. |
| Derivation |
AT3B-1 was derived from the AT-3 cell line. AT-3 was derived from a adult malignant rat prostate tumor obtained from a 22 month old inbred Copenhagen rat by Isaacs et al. When P-glycoprotein was blocked, the AT3 B-1 cell line demonstrated drug efflux pump activity. Injection of AT3 B-1 cells into rats followed by doxorubicin treatment produced larger tumors compared to the parental controls. |
| Comments |
AT3B-1 was derived from the AT-3 cell line. AT-3 was derived from a adult malignant rat prostate tumor obtained from a 22 month old inbred Copenhagen rat by Isaacs et al. The parental cells were grown in increasing concentrations of the drug doxorubicin until the final concentration of 0.001 mM was achieved. These cells express a multidrug (MDR) resistant phenotype. They are more resistant to vinblastine compared to controls. When P-glycoprotein was blocked, the AT3 B-1 cell line demonstrated drug efflux pump activity. Injection of AT3 B-1 cells into rats followed by doxorubicin treatment produced larger tumors compared to the parental controls. This cell line can be used to study chemotherapy resistance in prostate cancer. |
| 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-0.53mM 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 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: 1:3 to 1:8
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. |
| References |
Isaacs JT, et al. Establishment and characterization of seven Dunning rat prostatic cancer cell lines and their use in developing methods for predicting metastatic abilities of prostatic cancers. Prostate 9: 261-281, 1986. PubMed: 3774632
Replogle-Schwab TS, et al. Development of doxorubicin resistant rat prostate cancer cell lines. Anticancer Res. 17: 4535-4538, 1997. PubMed: 9494564
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.
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