266-6 (ATCC® CRL-2151)

Organism: Mus musculus, mouse  /  Tissue: pancreas  /  Disease: pancreatic acinar cell tumor

Organism Mus musculus, mouse
Tissue pancreas
Product Format frozen
Morphology epithelial
Culture Properties adherent
Biosafety Level 2  [Cells contain Papovavirus]
Disease pancreatic acinar cell tumor
Age adult
Applications The 266-6 cell line is useful for transfection studies.
Storage Conditions liquid nitrogen vapor phase
Derivation
266-6 is an acinar pancreatic cell line derived in 1985 by Robert E. Hammer from a young adult mouse.
The tumor was induced with an elastase I/SV-40 T antigen fusion gene.
Receptor Expression
acetylcholine, muscarinic
Comments
These cells retain a partially differentiated phenotype, and express detectable levels of a number of digestive enzyme mRNAs.
The cells respond to carbachol and cholecystokinin but not to substance P, secretin, or vasoactive intestinal peptide (VIP).
They bear an elastase I/neomycin transgene.
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%.
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.

  1. Remove and discard culture medium.
  2. 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.
  3. 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.
  4. Add 6.0 to 8.0 mL of complete growth medium and aspirate cells by gently pipetting.
  5. To remove trypsin-EDTA solution, transfer cell suspension to centrifuge tube and spin at approximately 125 x g for 5 to10 minutes.
  6. Discard supernatant and resuspend cells in fresh culture medium.  Add appropriate aliquots of cell suspension to new  0.1% gelatin coated culture  vessels (see Handling Procedure for Frozen Cells).
  7. Incubate cultures at 37°C.

Subculture Ratio: 1:3 to 1:4
Medium Renewal: 2 to 3 times a week.
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.

Cryopreservation
Freeze medium: Complete growth medium, 95%; DMSO, 5%
Storage temperature: liquid nitrogen vapor phase
Name of Depositor GH Swift
Year of Origin 1985
References

Kruse F, et al. The cell-specific elastase I enhancer comprises two domains [published erratum appears in Mol. Cell. Biol. 8: 1862, 1988]. Mol. Cell. Biol. 8: 893-902, 1988. PubMed: 3352608

Swift GH, et al. Differential requirements for cell-specific elastase I enhancer domains in transfected cells and transgenic mice. Genes Dev. 3: 687-696, 1989. PubMed: 2744460

Ornitz DM, et al. Elastase I promoter directs expression of human growth hormone and SV40 T antigen genes to pancreatic acinar cells in transgenic mice. Cold Spring Harbor Symp. Quant. Biol. 50: 399-409, 1985. PubMed: 3006998

Rose SD, et al. A single element of the elastase I enhancer is sufficient to direct transcription selectively to the pancreas and gut. Mol. Cell. Biol. 14: 2048-2057, 1994. PubMed: 8114736

Swift GH, et al. An element of the elastase I enhancer is an overlapping bipartite binding site activated by a heteromeric factor. J. Biol. Chem. 269: 12809-12815, 1994. PubMed: 8175694

Basic Documentation
References

Kruse F, et al. The cell-specific elastase I enhancer comprises two domains [published erratum appears in Mol. Cell. Biol. 8: 1862, 1988]. Mol. Cell. Biol. 8: 893-902, 1988. PubMed: 3352608

Swift GH, et al. Differential requirements for cell-specific elastase I enhancer domains in transfected cells and transgenic mice. Genes Dev. 3: 687-696, 1989. PubMed: 2744460

Ornitz DM, et al. Elastase I promoter directs expression of human growth hormone and SV40 T antigen genes to pancreatic acinar cells in transgenic mice. Cold Spring Harbor Symp. Quant. Biol. 50: 399-409, 1985. PubMed: 3006998

Rose SD, et al. A single element of the elastase I enhancer is sufficient to direct transcription selectively to the pancreas and gut. Mol. Cell. Biol. 14: 2048-2057, 1994. PubMed: 8114736

Swift GH, et al. An element of the elastase I enhancer is an overlapping bipartite binding site activated by a heteromeric factor. J. Biol. Chem. 269: 12809-12815, 1994. PubMed: 8175694