MATRIGEL Preparation:
(Basement Membrane Matrix 10 ml * LDEV-Free, BD 354234 – Store at -20 ºC)
- Thaw the Matrigel at 4 ºC throughout the night to avoid the formation of a gel. (The Matrigel canister should be placed in the refrigerator at 4ºC in a beaker containing water so that the defrosting process is gradual).
- The next day, add 10 ml of KO-DMEM medium to the 10 ml bottle of Matrigel and mix well with a sterile 10 ml pipette previously cooled.
- Aliquot in pre-refrigerated 15 ml tubes, adding 1 ml per tube with a sterile pipette, and put on ice.
- Store the aliquots at -20ºC.
1. Thaw the matrigel aliquot at least 10-15 min in a container with cold water at 4ºC.
2. Dilute each aliquot of 1 ml of Matrigel in MEM (dilution 1:15) and resuspend several times.
3. Add 25 ml of matrigel on the 500 cm2 dish and distribute well on the surface.
* It can be stored at 4ºC with parafilm for a maximum period of 1 week.
4. Before using the dishes, incubate at least one hour at room temperature and aspire the Matrigel before seeding the cells.