8
CORN I NG ASCENT FBR PD SYSTEM
Table 5-3.
Base, feed, and inoculum bottles descriptions.
Bottle
Description
Base (or alkali)
This bottle contains a basic solution to adjust cell culture media pH inside the MCV. The
Ascent system is designed to automatically maintain the MCV cell culture media pH by
adding base solution via the base pump and controlling pump flow with user-defined
setpoints. Adding a base solution during cell growth neutralizes excessive concentration of
H+ ions and hence prevents the media from being too acidic. An example of a base solution is
7.5% sodium bicarbonate solution.
Feed (or bolus)
This bottle contains concentrated supplement(s) that, when added to the media, can increase
nutrient concentrations during cell culture. Feed is pumped into the MCV at a programmed
rate via the feed pump.
As an option, nutrients such as glucose (45%) and L-glutamine (200 mm) can be added in
bulk via the feed line to increase concentrations.
Inoculum (or cell
suspension)
This bottle contains a concentrated cell suspension in media that is used to seed the
bioreactor. Seeding can be done through the MCV.
5.1.5 FBR with Sterile Connectors
The Corning® Ascent™ FBR system has two options to connect media flow from the MCV to the FBR and return media from the FBR to MCV.
Option A uses Lynx® connectors while Option B uses AseptiQuik® G connectors (Figure 5-6 and Figure 5-7). At experiment start, the sterile
connectors (Option A or B), are connected to their mating sterile connectors on the MCV.
The Lynx® connectors provide users the added benefit of aseptically connecting and disconnecting the FBR up to six times for manual
manipulations enabling substrate sampling. The portable tray allows users the convenience of moving the disconnected FBR and Lynx®
connectors to a biosafety cabinet. The AseptiQuik® G is a single-use connector and cannot be disconnected or reconnected.
5.1.4 Inoculum, Base, and Feed Bottles
Inoculum is manually added during an experiment. The user selects one of two AseptiQuik® G connectors on the MCV headplate (see
Figure 5-2). Base is automatically added as part of the pH control loop. Base and feed lines have respective pumps that require set up prior
to use. Figure 5-5 shows a rack for base, feed, and inoculum bottles. Refer to Table 5-3 for descriptions.
Figure 5-5.
Base and feed bottles are connected to their respective
pumps during an auto-cycle. Inoculum is manually introduced.