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Isolation:
Isolation date: May 5, 1995
Tumorigenic:
Yes
Oncogene:
K-ras +
Antigen Expression:
MHC class I +; MHC class II - [50655]
Blood type B; Rh+
Age:
77 years *****
Gender:
female
Ethnicity:
White
Comments:
Panc 05.04 is a pancreatic adenocarcinoma epithelial cell line derived, in 1995, from a primary tumor removed from the head-of-the-pancreas of a female with pancreatic adenocarcinoma.
The cell line exhibits a K-ras oncogene mutation at codon 12 where a GGT --> GAT mutation resulted in substitution of aspartic acid for glycine. [50655]
The cells have a reported plating efficiency of 100%. [50655]
Propagation:
ATCC complete growth medium: RPMI 1640 medium with 2 mM L-glutamine adjusted to contain 1.5 g/L sodium bicarbonate, 4.5 g/L glucose, 10 mM HEPES, and 1.0 mM sodium pyruvate and supplemented with 20 Units/ml human recombinant insulin, 85%; fetal bovine serum, 15% Atmosphere: air, 95%; carbon dioxide (CO2), 5% Temperature: 37.0°C
Subculturing:
Protocol:
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 centrifuge tube and spin at approximately 125 xg 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.
Subcultivation Ratio: A subcultivation ratio of 1:3 to 1:5 is recommended Medium Renewal: Add media once per week. Fluid change one to two times per week.
Preservation:
Freeze medium: Complete growth medium supplemented with 5% (v/v) DMSO Storage temperature: liquid nitrogen vapor phase
Doubling Time:
46 hrs
Related Products:
Recommended medium (without the additional supplements or serum described under ATCC Medium):ATCC 30-2001
recommended serum:ATCC 30-2020
References:
50655: Jaffee EM, et al. Development and characterization of a cytokine-secreting pancreatic adenocarcinoma vaccine from primary tumors for use in clinical trials. Cancer J. Sci. Am. 4: 194-203, 1998. PubMed: 9612602
