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This cell line can be paired with MEF cell lines, generated from BCL-2 family member knockout mice (e.g., ATCC CRL-2908, ATCC CRL-2909, ATCC CRL-2910, ATCC CRL-2911, ATCC CRL-2912, ATCC CRL-2913), as a control to study the molecular mechnaisms of cell apoptosis and the BCL-2 family signaling pathway.

These cells are SV40 immortalized mouse embryonic fibroblasts (MEFs), which were generated from wild type mice.

These cells are SV40 immortalized mouse embryonic fibroblasts (MEFs), which were generated from wild type mice.

This cell line can be paired with MEF cell lines, generated from BCL-2 family member knockout mice (e.g., ATCC CRL-2908, ATCC CRL-2909, ATCC CRL-2910, ATCC CRL-2911, ATCC CRL-2912, ATCC CRL-2913), as a control to study the molecular mechnaisms of cell apoptosis and the BCL-2 family signaling pathway.

The cells will slough off of the growth surface if they become too heavy.

1. Remove and discard culture medium.
2. Briefly rinse the cell layer with Ca++/Mg++ free Dulbecco's phosphate-buffered saline (D-PBS) or 0.25% (w/v) Trypsin - 0.53 mM EDTA solution to remove all traces of serum which contains trypsin inhibitor.
3. Add 1.0 to 2.0 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 37.0°C to facilitate dispersal.
4. Add 6.0 to 8.0 mL of complete growth medium and aspirate cells by gently pipetting.
5. Add appropriate aliquots of the cell suspension to new culture vessels. An inoculum of 5 X 10³ to 1 X 10⁴ viable cells/cm² is recommended.
6. Incubate cultures at 37°C. At subculture, cell concentration is between 1 X 10⁵ to 2 X 10⁵ cells/cm².

Wei M, et al. Proapoptotic BAX and BAK: a requisite gateway to mitochondrial dysfunction and death. Science 292(5517): 727-730, 2001 PubMed: 11326099

Lindsten T, et al. The Combined Functions of Proapoptotic Bcl-2 Family Members Bak and Bax Are Essential for Normal Development of Multiple Tissues. Molecular Cell 6(6): 1389-1399, 2000 PubMed: 11163212

Zong WX, et al. BH3-only proteins that bind pro-survival Bcl-2 family members fail to induce apoptosis in the absence of Bax and Bak. Genes Dev. 15(12): 1481-6, 2001 PubMed: 11410528

Cheng EH, et al. BCL-2, BCL-XL Sequester BH3 Domain-Only Molecules Preventing BAX- and BAK-Mediated Mitochondrial Apoptosis. Molecular Cell 8(3) : 705–711, 2001 PubMed: 11583631