Team:Freiburg/Protocols/Freeze Thaw

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Freezing and Thawing Cells

Freezing and Thawing Cells

material: DMSO, FCS, cells, cryotubes…
duration: 20 min

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1. prepare your freeezing Medium (50& FCS, 40% Medium, 10% DMSO; may vary deending on your cell line) and cool it
2. superscripe the Cryotube (Celltype, passage number, amount of cells, Date,..)(use some Tesa to stick over your writing)
3. harvest the cells
4. spin down cells (100-300 x g, 5 min)
5. resuspend cell pellet in 3 ml cold Cryomedium and fill 1,5 ml into each Cryotube (DMSO is harmfull for cells, so time is of essence)
6. put the tubes in a freezing box filled with isopropanol and put the whole thing in the -80 ° C freezer.
7. After at least 3 h you can transfer the cryotubes to their long-term stock

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