3.15 Growing
M. tuberculosis Using
a Fermentor/
Bioreactor Under
Normoxic and Hypoxic
Conditions
See Fig. 11 for the bioreactor setup.
1. Add media to the bioreactor using a peristaltic pump. This will
involve setting up a filtration system to insure media sterility.
Place an empty reactor vessel, peristaltic pump, and the media
into the BSC with the pump in the middle.
2. Remove the lid of the vessel to expose the tubes. Make sure the
exhaust line does not have any kinks that may prevent proper
flow of the media. Locate and unwind one of the large liquid
addition (LA) ports. Clamp the LA line closed and cut off the
tip of the line, removing the tube attachment adaptor. Fit the
open end of the LA tube to the outlet port of a VacuCap
located under the VacuCap bottle fitting. Use a clamp to secure
the tube tightly.
3. Attach the feeder line of the VacuCap through the peristaltic
pump. Remove the buoy from the end of the feeder line.
4. Remove cotton from a 10 mL serological pipet (or a 10 mL
aspiration pipette). Insert the pipette into the feeder line to
create a media harvesting wand. Insert the wand into the media
flasks and turn on the pump at max speed. The media will be
pulled from the flask, filtered through the VacuCap, and dispensed into the sterile vessel.
Fig. 11 Bioreactor setup for hypoxic and normoxic paired growth
46
Elizabeth Wallace et al.