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PL45 is a pancreatic adenocarcinoma epithelial cell line derived in 1992 from a primary tumor removed from the pancreas of a man with poorly differentiated pancreatic adenocarcinoma of ductal origin. The MTS1 gene on chromosome 9p21 encodes the p16 inhibitor of cyclinD/CDK4 complexes. PL45 cells have a wild-type p16 gene. [50656] This gene is transcriptionally silenced and methylated in PL45 cells. PL45 cells have a wild-type cyclin-dependent kinase (CDK4) gene. Both the PL45 and the Panc 10.05 cell lines exhibit a K-ras oncogene mutation at codon 12 where a GGT --> GAT mutation resulted in substitution of aspartic acid for glycine. [50655] [50656] The line expresses wild-type DPC4 and BRCA2 genes. The line has a p53 gene mutation at codon 255 where an ATC --> AAC mutation resulted in substitution of asparagine for isoleucine. The PL45 cell line was derived from the same patient as the Panc 10.05 cell line (ATCC CRL-2547).
Propagation:
ATCC complete growth medium: The base medium for this cell line is ATCC-formulated Dulbecco's Modified Eagle's Medium, Catalog No. 30-2002. To make the complete growth medium, add the following components to the base medium: fetal bovine serum to a final concentration of 10%. 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.
Add appropriate aliquots of the cell suspension to new culture vessels.
Incubate cultures at 37°C.
Subcultivation Ratio: A subcultivation ratio of 1:2 to 1:6 is recommended Medium Renewal: Every 2 to 3 days
Preservation:
Freeze medium: Complete growth medium supplemented with 5% (v/v) DMSO Storage temperature: liquid nitrogen vapor phase
Related Products:
Recommended medium (without the additional supplements or serum described under ATCC Medium):ATCC 30-2002 recommended serum:ATCC 30-2020 derived from same individual:ATCC CRL-2547
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 50656: Caldas C, et al. Frequent somatic mutations and homozygous deletions of the p16 (MTS1) gene in pancreatic adenocarcinoma. Nat. Genet. 8: 27-32, 1994. PubMed: 7726912 51521: Su GH, et al. ACVR1B (ALK4, activin receptor type 1B) gene mutations in pancreatic carcinoma. Proc. Natl. Acad. Sci. USA 98: 3254-3257, 2001. PubMed: 11248065
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