Applications |
DSL-6B/C2 is a pancreatic ductal cell line derived from the DSL-6 transplantable acinar cell carcinoma. The DSL-6 tumor was established in 1986 from a primary acinar cell carcinoma of the pancreas which developed in a male Lewis rat (DSL-101-79) that was given azaserine intraperitoneally. The cultured DSL-6B/C2 tumor cells initially produced amylase, but production of exocrine enzymes ceased after 1 to 2 weeks in culture. |
Derivation |
DSL-6B/C2 is a pancreatic ductal cell line derived from the DSL-6 transplantable acinar cell carcinoma. The DSL-6 tumor was established in 1986 from a primary acinar cell carcinoma of the pancreas which developed in a male Lewis rat (DSL-101-79) that was given azaserine intraperitoneally. The cultured DSL-6B/C2 tumor cells initially produced amylase, but production of exocrine enzymes ceased after 1 to 2 weeks in culture. |
Comments |
DSL-6B/C2 is a pancreatic ductal cell line derived from the DSL-6 transplantable acinar cell carcinoma. The DSL-6 tumor was established in 1986 from a primary acinar cell carcinoma of the pancreas which developed in a male Lewis rat (DSL-101-79) that was given azaserine intraperitoneally. The cultured DSL-6B/C2 tumor cells initially produced amylase, but production of exocrine enzymes ceased after 1 to 2 weeks in culture. The cell line also lost structural and immunohistochemical acinar cell markers while acquiring duct cell markers during culture and regrafting. |
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 Ca++/Mg++ free Dulbecco's phosphate-buffered saline (D-PBS) or 0.25% (w/v) Trypsin-0.53 mM ) 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 1:4
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 |
Durfee T, et al. Derivation of ductlike cell lines from a transplantable acinar cell carcinoma of the rat pancreas. Am. J. Pathol. 143: 292-303, 1993. PubMed: 8391218
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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