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We’ve got it in the bag.
Multitron II Bag Configuration
The Multitron with Shaker Bag option enables the cultivation of mammalian, insect or plant cells in disposable bags from 1 to 10 L working volumes. Direct gassing of air or gas mix of air/CO2 provides aeration and pH stability for each individual bag. The Multitron Bag Shaker is designed to accommodate standard bags from any manufacturer.
Configurations are available for one large disposable bag or multiple small disposable bags. The strategic placement of a PT100 temperature sensor provides accurate temperature monitoring and control. Rotameters are conveniently located on the side of the unit to control gas flow rate. Alternatively rotameters can be mounted inside the cabinet to reduce total width.
A specially designed passage on the side of the incubator allows cables and gassing lines to be easily led into the cultivation chamber.
New Option for Incubator Shaker: Cell cultivation with orbital waves in flexible single-use shaker bags up to 10 L
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.
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.
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.
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