| MEF DR4 irradiated |
|
| Catalog # | E2256-16 |
| Cell Type | Mouse Embryonic fibroblasts |
| Description | Mouse embryonic fibroblast cells, resistant to neomycin, hygromycin, puromycin and 6-thioguanine. Passage #3, irradiated, ~2x106 cells/vial |
| Note | All cell lines are produced under aseptic conditions. Each lot is tested to be free from bacterial/fungal, mouse pathogen and mycoplasma contamination before shipping. |
| ANALYSIS PARAMETERS CRITERION RESULT Viability Test1 > 60% 95% Sterility Test2 no growth no growth Mouse Pathogen Test3 negative negative Mycoplasma4 negative negative Mouse ESC support5 pass pass |
| 1. Viability is defined as the ratio of number of live cells over total number of cells after thawing. 2. Immersion method is used to inoculate directly into Tryptic Soy Broth media and Fluid Thioglycollate media to check for bacteria and fungal contamination. 3. PCR based assays for mouse pathogens including Sendai virus, mouse hepatitis virus, pneumonia virus, minute virus, parvovirus (MPV1,MPV2, MPV3) and Theiler’s murine encephalomyelitis virus. 4. PCR based assay. 5. Mouse embryonic stem cells are plated and observed for growth on a layer of feeder cells by ES cell colony morphology. |
| Recommended Procedure | Preparing MEF DR4 as feeder cells: |
| 1. Place a vial of frozen cells at 37 ºC water bath immediately after removing from liquid nitrogen freezer. As soon as contents of the vial start thawing, transfer contents to a new tube containing 10x volume of prewarmed growth medium. |
| 2. Spin at 1000 rpm for 5min, discard medium, resuspend cells in growth medium and plate cells in two 100mm culture dishes. |
| 3. On the second day, change the medium with ES culture medium, and they are now ready to use as feeder for ES cells. |
| Suggested Drug Concentration | 200ug/ml for neomycin (G418), 1ug/ml for puromycin, 120ug/ml for hygromycin, and 1ug/ml or 6-thioguanine. |
| Storage | Vapor phase of liquid nitrogen freezer. |
| Important Note | This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications without the expressed written authorization of United States Biological. |
