| Applications |
TGP52 is an epithelial like cell line derived from a pancreatic tumor with islet cell morphology (insulinoma) arising in an adult male transgenic mouse. The mouse carried the pseudogene construct composed of elastase-1 promoter linked to the SV40 T antigen. In culture, TGP52 cells transiently produced insulin in culture and then produced somatostatin. Current stocks of TGP52 secrete about 9700 pg/ml of somatostatin in 3 days. |
| Comments |
TGP52 is an epithelial like cell line derived from a pancreatic tumor with islet cell morphology (insulinoma) arising in an adult male transgenic mouse. The mouse carried the pseudogene construct composed of elastase-1 promoter linked to the SV40 T antigen. In culture, TGP52 cells transiently produced insulin in culture and then produced somatostatin. Current stocks of TGP52 secrete about 9700 pg/ml of somatostatin in 3 days. Morphologic and differentiation markers of this line appear to fluctuate from time to time in culture. |
| 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 1:4
Medium Renewal: Every 2 to 3 days
Note: For more information on enzymatic dissociation and subculturing of cell lines consult Chapter 13 in Culture of Animal Cells, a Manual of Basic Technique by R. Ian Freshney, 5th edition, published by Wiley-Liss, N.Y., 2005. |
| References |
Pettengill OS, et al. Cell lines derived from pancreatic tumors of Tg(Ela-1-SV40E)Bri18 transgenic mice express somatostatin and T antigen. Carcinogenesis 15: 61-65, 1994. PubMed: 7904904
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.
|
