The newest addition to our line of highly flexible incubated shakers from

• Unbeatable flexibility:
   - compatible with microplates, flasks, or bags
• Stackable, space efficient
• Controlled uniform temperature
• Contained growth environment
• Gas delivered via controlled environmental air pump
• Air and CO2 delivered via rotameter to each bag
• Compatible with any manufacturer’s bags
• Bag capacity per shaker:
   - 3 x 2L (1L Vw), 2 x 10L (5L Vw), or 1 x 20L (10L Vw)

Specifications

• For mammalian and insect cell cultivation - Suitable for all types of eucaryotic cells
• Uses standard disposable bags from any manufacturer
• Incubation Chamber - Disposable bioreactor bags are incubated in the Multitron chamber and heated to the precise working temperature. Tubing and filters are also contained in the shaker allowing for more uniformity. Heated filters help to reduce condensation and filter blockage.
• Gassing - For insect cells air from within the Multitron is pumped through the disposable bioreactor bag. For mammalian cells the Multitron is equipped with an IR sensor to accurately control CO2. The chamber CO2 enriched mixture is pumped through the disposable bioreactor bag.
  The gas flow rates are precisely controlled with rotameters.
• Excellent temperature control - A mobile PT100 measures liquid temperature directly at the bag, providing precise and even temperature control.
• Different sizes - 3 x 2 L, 2 x 10 L or 1 x 20 L total volume disposable bags can be incubated in the Multitron.
• Pre-defined packages - Simple, pre-defined packages make specification and ordering easy.
• Unbeatable flexibility - The shaker bag tray can be easily exchanged for universal trays with Sticky Squares or clamps. Simply slide the shaker bag tray out and slide another tray in. The result is the most flexible platform for growing cells of all types.

Application Note

New Option for Incubator Shaker: Cell cultivation with orbital waves in flexible single-use shaker bags up to 10 L

Introduction

Orbitally shaken, single-use bags represent a new method for the production of seed cultures or protein production on the lab scale.

Using this new technology, successful cultivation of the cell lines CHO XM 111 (Chinese Hamster Ovary) and Sf-21 (Sf-9) (Spodoptera frugipedra) was accomplished. Also, scale-up of the CHO cell cultivation from 1 L to 10 L working volume is possible using orbital-shaken, single-use bags. For these cell cultures a serum- and protein-free chemically defined media was used, which needs sophisticated system efforts for the cell growth, but is a requirement for the production of therapeutic products.

Details

The CHO and Sf-9 cell lines are frequently used in biotechnology. For this experimental work, the CHO XM 111 clone was used. This clone was transformed by the group of Professor Dr. M. Fussenegger at the ETH Zurich with an expression vector which codes for the gene of the recombinant protein SEAP (Secreted Alkaline Phosphatase) and controlled by tetracycline using the promoter Ph- CMV-1. The Sf-9 is a cell clone of the cell line Sf-21. These insect cells, developed from ovaries of the moth species Spodoptera frugipedra, are well known for virus infection and production of recombinant proteins.

The Multitron Cell for shaker bags was used with a 50 mm shaker stroke and aeration with maximal 2 vvm air and CO2 (0–10% only for CHO cells). The cultivation of CHO XM 111 were done at 37°C, pH 7.2 and shaker speed between 30 and 40 rpm. For the cultivation of Sf-9 cells a temperature of 27°C, shaker speed between 25 and 35 rpm and pH 6 were used.

The oxygen transfer was determined by amperometric method.

Result

The fed-batch cultivation strategy was used in chemical defined media:

• CHO (protein- and serum-free): HP-1 (Cell Culture Technologies, Invitrogen) with 2.5 mL L–1 Tetracyclin and 10mL L–1 Pluronic F-68

• Sf-9 (serum-free): Sf-900 II SFM (Cell Culture Technologies) Cell growth results in maximal cell densities for

• CHO XM 111 about 3x106 viable cells/ml with > 98% viability

• Sf-21 (Sf-9) > 1x107 viable cells/ml with > 90% viability. An Oxygen Transfer Coefficient (kLa) about 31 h–1 with 2 vvm could be achieved.