Cell Transient Transfection Workstation

Automated small scale transient transfection system

This application note describes an automated small scale transient transfection system. Cells have to be transfected with different type of DNA constructs in order to produce specific proteins. The cell culture is divided from a 3L spinner flask to 48 CultiFlasks. These CultiFlasks were stored for several days in an incubator.

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System Requirements
All open containers/tubes etc. have to be handled in a sterile environment (product protection, safety class 1). A complete CultiFlask (meaning pipetted DNA, Transfection reagent and cells together) should be no longer than 1hr on the system.

Process:
Day 1. Transient Transfection
Two different DNA constructs and transfection reagent were pipetted in media into a CultiFlask. After short incubation cell suspension is added and the CultiFlask will be stored in an incubator overnight.

Day 2. Adding Feeding Solutions
Three different feeding solutions were added to the cell suspension into the CultiFlask. The CultiFlask will be stored in an incubator for 4 days.

Day 5. Measuring Yield
From each CultiFlask a sample is taken and added to an 96w MTP. If the yield is enough the CultiFlasks will be harvested by centrifugation (300g) and a filtration step 0.22µm, all steps automatically  performed by Synchrons’ Robotic platform.

Synchron Application
• Synchron Xperimate x-y-z robotic platform
• 2.50 meter system with 2.0 meter deck and 0.50 meter overhang for getting access to robotic centrifuge
• Robotic arm with 4 channels, 2 channels equipped with  SX48 micro-annular gear pumps, 2 others equipped with SX3 micro- annular gear pumps
• 2 channels equipped with custom made pipetting tips so that they can pipette directly onto 0.22µm Luor-Lock filters, 2 others equipped with disposable tips
• Flexispan
• Chemical resistant tubing, valves and pumps
• 2nd robotic arm with tube gripping tool
• Fully integrated 2D Tube scanner by Ziath
• Fully integrated Shaker equipped with 48 position tube rack
• Fully integrated De-/Capping module
• Fully integrated dispensing module for adding cell suspension
• Fully integrated robotic centrifuge by Hettich
• Magnetic stirrer for Spinnerflask by IKA
• Peristaltic pump
• Customized rack for 24 positions 0.22µm Luor-Lock filters
• Customized racks for 24 positions 50mL Tubes