Steps for using a multi-layer cell culture vessel:
For 10-layer and 40-layer chambers, pre-warming the chamber and medium is especially important due to the increased time required for the middle chamber to reach the desired incubation temperature.
Unpack the chamber and place the chamber inside a fume hood or cleanroom environment to make aseptic connections.
Replace one vent cap with a vent filter assembly. Connect the cell suspension tubing to another port of the chamber. Place the chamber with the air filter facing up. Open the clamp to begin filling. The cell suspension will spread evenly between the trays during filling.
Once the cell suspension has been evenly distributed in the previous position, turn the chamber upside down so that the ports are facing up.
Close the clamp to seal the tube or heat seal the tube or replace the filling tube with a vent filter assembly cap or vent cap.
Gently lower the chamber to its normal horizontal incubation position and gently tilt the chamber back and forth until the surface of each chamber is completely covered with medium. This will ensure even distribution of cells over each growth surface.
Place the chamber in an incubator or warm room. Due to the pressure build-up during incubation, the chamber must be vented during incubation.
To empty the chamber, replace one of the vent caps with a drain. Tilt the chamber on its side with the air filter facing up. Make sure the chamber is above a sterile collection vessel if emptying by gravity. Tilt the chamber to drain
