11. Attach a 5 cm tubing to the valve and a Tee connector to

it. Then,

(a)

Attach two 5 cm silicone tubing to the Tee connector.

(b)

Connect a one-way valve to one tubing (flow direction

towards the tee connector) and a 0.2 μm filter to it.

(c)

Attach another one-way valve to the second tubing and

connect a 5 cm tubing to its other end. Connect a 60 mL

syringe to it, this syringe will be positioned later onto the

syringe pump.

12. Attach a 15 cm tubing (Tube-A) to the PBS long tubing tee

connector.

(a)

Connect it to one SDS tubing using another tee

connector.

(b)

Attach a 15 cm tubing to the third end of the tee connec-

tor. This tubing will be connected to the bioreactor

LEFT side.

13. Attach a 15 cm tubing to the second SDS tubing. Connect it

with a Tygon^® 3-stop tubing using a fitting reducer.

14. Attach a 15 cm tubing to the Tygon^® using a fitting reducer.

Connect the tubing to another 15 cm tubing using a tee

connector. This tubing will be connected on the RIGHT side

of the bioreactor.

15. Attach a 15 cm tubing (Tube-B) to the last end of the tee

connector and insert it into the waste bottle.

16. Put aside the flasks and tubing circuitries, while keeping them

in the BSC.

Fig. 2 Schematic representation of the bioreactor system setup for tracheal de-epithelialization. Length of

certain tubing is specified on the side in centimeters. Tube-A and Tube-B connect the SDS circuitry with the

detergent-waste system. Electronic circuitry not shown

174

Alba E. Marin-Araujo et al.