| 產品名稱 |
266-6 |
| 商品貨號 |
B163653 |
| Organism |
Mus musculus, mouse |
| Tissue |
pancreas |
| Product Format |
frozen |
| Morphology |
epithelial |
| Culture Properties |
adherent |
| Biosafety Level |
2 [Cells contain Papovavirus]
Biosafety classification is based on U.S. Public Health Service Guidelines, it is the responsibility of the customer to ensure that their facilities comply with biosafety regulations for their own country. |
| 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.
Cells must be seeded on 0.1% gelatin-coated culture vessels. To make the coating, dissolve 1.0 g of Gelatin Type A (Sigma G2500) in 1L of distilled water, sterilize by autoclaving. Add 0.4 to 0.5 mL of gelatin solution per cm2 of the culture vessel and refrigerate for 10 to 15 minutes. Prior to use remove the gelatin solution and rinse gently with culture medium, adding at least 0.1 to 0.2 ml per cm2. Discard the culture medium used for rinsing and use vessels immediately. If required, coated vessels can be prepared in advance and stored at 2 to 8?C no more than 5 days.
- 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 x g for 5 to10 minutes.
- 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).
- 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 |
| Deposited As |
Mus musculus |
| 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
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