Passaging of C2C12 cells

Passaging of C2C12 cells

Making a New Passage C2C12 cells

Summary

We passage cells to keep them alive and growing in the incubator in the cell culture room. Generally cells should be passaged every three days

Materials

• Cell plates
• Proliferation media
• PBS
• Trypsin

Methods

1. Warm all solutions needed (PBS, prolif media) trypsin is not warmed
2. Label new dish (cell type/ passage number/ date/ initials/ prolif)
3. Add 10 ml of prolif media to new plate
4. Take up old cell media with vacuum and pipette tips
5. Wash old plate with 10 ml of PBS and remove
6. Add 1 ml of trypsin and place in incubator for two minutes (or until cells are detached)
7. Once cells are detached flush old plate a few times and take up 100 ul of cells (fridays- 70 ul) and drop five drops onto the dish
8. Flush new dish with cells a few times
9. Place in incubator, gently shake plate in incubator to spread cells and media