3.3.2

Bioreactor Setup

1. Open the sterile tray package inside the BSC (see Note 8).

Then, open the autoclaved main and extra bioreactor compo-

nents, and place them on the tray.

2. Position the motor bracket onto the bioreactor anterior wall.

Place four washers between the bracket and the bioreactor.

Using an Allen key, screw on the bracket with four screws

(Fig. 1a).

3. Insert a barbed tube fitting adapter into the short anchor

threaded end, and a swivel joint into the long anchor threaded

end (see Note 9).

4. Place one O-ring on each end of the bioreactor body. Lubricate

the O-rings with the autoclaved silicone-based lubricant using

a sterile swab/applicator.

5. Place the two rotating anchors on opposite ends of the bior-

eactor’s body; the anchors should be in contact with the

O-rings. Place the short anchor on the left end and the long

one on the right end.

6. Adjust the position of the long anchor with respect to the

bioreactor body to accommodate the trachea (note that the

system would allow a trachea of any length from 3 to 10 cm).

Insert the large pulley into the long rotating anchor, secure it

by tightening the screw.

7. Wrap the thread portions of two plastic barbed tube fittings

with Teflon tape to ensure water-tight threads. Insert one

barbed tube fitting into the threaded hole at the anterior

bioreactor wall, and another tube fitting at the posterior one.

8. Wrap the temperature sensor threaded section with Teflon

tape. Screw the sensor long probe into the second hole at the

bioreactor anterior wall.

9. Using the pliers, make sure the fittings and the sensor have a

tight seal.

10. Connect a 5 cm silicone tube to each barbed tube fitting

protruding from the bioreactor body (anterior and posterior

walls). Place one plastic cap into each previously positioned

tubing.

11. Introduce the 15 cm tubing (Subheading 3.3.1, step 15, i.e.,

bioreactor RIGHT side tubing) into the shaft placed in the

shaft holder; secure the tubing by tightening the shaft screw.

Place the shaft holder approximately 6–7 cm from the bioreac-

tor using its rod; secure the shaft holder by tightening its screw

at the bottom onto the rod. Attach the rod-shaft holder to the

right side of the bioreactor; secure its position tightening the

screw at the bottom of the bioreactor body.

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