Passaging C2C12 cells

Materials:

• 1x 15ml FALCON tube
• PBS w/o Ca2+ and Mg2+
• TE in PBS (4 ml TE + 36 ml PBS)
• HS for C2C12 cells
• pipettes
• 75 cm2 flask

procedure:

• remove 5 ml of the old media in the 15 ml FALCON tube
• throw the rest of the old media away

- wash the cells with 5 ml PBS - remove the PBS - add 2 ml of the TE in the dish - let this incubate for ~4min at 37ºC - stop the TE with the old media and resuspend the cells in this - pipette the cell suspension in the FALCON tube back - centrifuge the suspension for 1000 xg for 3 min at 4 ºC - remove the old media - resuspend the cells in an appropriate volume of HS medium (f.e.: 10ml) - fill in the new 75 cm2 flask 14 ml of new media - pipette an appropriate volume of the cell suspension in he new flask and resuspend it in the new flask (f.e.: if you want to dilute your cell 1: 20 put in the new flask 0.5 ml of the resuspended cells in the FALCON tube) - close the flask and put it in the 37 ºC