4. Assemble pH sensor, dissolved oxygen (DO) sensor, all

stainless-steel tubes and impellers to the respective ports on

the top plate (Fig. 3), according to manufacturer’s instructions.

5. Fill the culture vessel with 2 L PBS and assemble the top plate

to the culture vessel and tighten the bolts to ensure the vessel is

airtight.

6. Remove the accessories from accessory bags and plug the sili-

con tubes 1–8 to their respective ports on the top plate (see

Note 4).

7. Preform a pressure test to ensure the vessel is airtight (see Notes

5 and 6).

8. Wrap all luer connectors with aluminum foil and tighten all

Robert clamps on the silicon tubes 1–6. Leave tube 8 open for

pressure relieve during autoclave (Fig. 3, see Note 7).

9. Autoclave the assembled culture vessel at 121 ^C for 60 min

(see Note 8). Remove from the autoclave and cool to room

temperature. Remove all hemostatic forceps if used.

10. Install the rotor motor and connect to the controller as accord-

ing to manufacturer’s instructions. Insert the temperature

probe and wrap the heating mat around the culture vessel.

Connect compressed air and CO2 to the controller. Fit the

silicon tubes 1 and 2 to pump 1 and 2, respectively (Fig. 4,

see Note 9).

11. Unclamp the Robert clamps on tubes 5, 6, and 8. On the

controller, set temperature to 37^C, agitation speed to

100 rpm/min and inlet air to 22 ccm.

Fig. 3 Ports on the top plate of 3D FloTrix^® vivaSPIN bioreactor and their respective functions

Large-Scale Expansion of UCMSCs with Microcarrier Tablets

117