Description

Diabetic Mouse Colonic Epithelial Cells from Cell Biologics are isolated from the colon of Diabetic (db/db) mice. Diabetic Mouse Colonic Epithelial Cells are grown in T25 tissue culture flasks pre-coated with gelatin-based coating solution for 2 min and incubated in Cell Biologics’ Culture Complete Growth Medium generally for 3-7 days. Cultures are then expanded. Prior to shipping, cells at passage 3 are detached from flasks and immediately cryo-preserved in vials. Each vial contains at least 0.5x10⁶ cells per ml. The method we use to isolate primary epithelial cells was developed based on a combination of established and our proprietary methods. Cells are incubated with EpCAM-1 (CD326) antibody, followed by the application of magnetic beads pre-coated with secondary antibody.

Product Testing

Diabetic Mouse Colonic Epithelial Cells are characterized by immunofluorescent staining with antibodies of E-cadherin (Catalog 610182, BD), ZO-1 (Catalog 617300, Life Technology). Cells are negative for bacteria, yeast, fungi, and mycoplasma. Cells can be expanded for 3-7 passages at a split ratio of 1:2 under the cell culture conditions specified by Cell Biologics. Repeated freezing and thawing of cells is not recommended.

Suggested Medium

Epithelial Cell Medium /w Kit –500 ml

Catalog No. M6621

Laboratory Applications

Diabetic Mouse Colonic Epithelial Cells can be used for the assay of cell-cell interaction, adhesion, PCR, Western blot, immunoprecipitation, immunofluorescent flow cytometry, or generating cell derivatives for desired research applications.

Storage

Cell Biologics ships frozen cells on dry ice. On receipt, immediately transfer frozen cells to liquid nitrogen (-180 °C) until ready for experimental use. Live cell shipment is also available on request. Primary cells should never be kept at -20 °C.

Documents

MD-6047

Cell-Culture-Protocol-CB

Disclaimer

Appropriate safety procedures should always be used with this material. Investigators should handle the cells that they receive from Cell Biologics with caution and treat all primary cells as potential pathogens, since no test procedure can completely guarantee the absence of infectious agents. The entire text of discussing Biosafety in Microbiological and Biomedical Laboratories, 5th ed. is available online at http://www.cdc.gov/biosafety/publications/bmbl5/index.htm.