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Hybridoma Cell Culture Application Using a BioFlo 110 Benchtop Bioreactor


Introduction
This Application Report is one in a series documenting simple protocols for culturing cells in a BioFlo 110 benchtop fermentor/bioreactor system. The modular BioFlo 110 is offered with an extensive choice of vessels, impellers and control modules to enable growth of a wide variety of cultures, including mammalian, insect, plant, yeast and bacterial cells. Complete kits are also available. This report concentrates on achieving highdensity hybridoma cell cultures using NBS Advanced Cell Culture Kit.

For this application, researchers in NBS cell culture laboratory used a standard 3.0L BioFlo 110 Advanced Cell Culture Kit with a magnetic drive and a water-jacketed vessel (NBS Catalog # M1273-1212). A suspension cell spin filter, with a 12-14 micron screen (NBS Catalog # M1273-3202) was used for perfusion in order to grow the hybridoma cells in a continuous mode. Additionally, BioCommand Plus supervisory software (NBS Catalog # M1291-0000) was used to control the feeding schedule and collect data; and a NucleoCounter (NBS Catalog # M1293-0000) was used to count viable cells.


The Bioreactor

Vessel
This BioFlo 110 Advanced Cell Culture Kit was equipped with a magnetic drive and water-jacketed 3.0 L vessel with a nominal working volume of 2.25 L. All BioFlo 110 Advanced Cell Culture Kits are configured with six control modules, a complete vessel assembly including a pitched blade agitation impeller, foam/level sensors, as well as dissolved oxygen and pH probes (Mettler Toledo), three liquid addition bottles, cables, tubing, clamps and other related items. Optional direct drive and magnetic-drive agitation systems, as well heat-blanketed vessel options, are available.

For this application
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