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•Development Of An Animal-Component Free Electroporation And Recovery Formulation Using EX-CELL™ CHO Cloning Medium

Electroporation is widely used for gene delivery in cell line generation, and is the method of choice for serum-free suspension culture transfection. Electroporation protocols and media can have a major impact on post-electroporation cell viability and transfection efficiency. Many laboratories use Phosphate Buffered Saline (PBS), various buffers or growth media for electroporation, which can lead to variable post-electroporation viabilities and transfection levels. We have optimized a single electroporation and recovery medium in an effort to increase post-electroporation cell viability, recovery viable cell density, and protein expression levels for Chinese Hamster Ovary (CHO) Cells.

We demonstrated that EX-CELL CHO Cloning Medium (C6366) supports electroporation of plasmid DNA as well as siRNA into CHO K1 cells. We conducted electroporation and recovery media optimization using Design of Experiments (DOE) by mixing EX-CELL CHO Cloning Medium (C6366) with two other SAFC Biosciences media in order to further improve transfection effi ciency and post-electroporation recovery. By optimizing post-electroporation viabilities, viable cell densities and protein expression levels, we were able to create an optimized formulation. This optimized formulation achieved equivalent or higher post-electroporation viabilities, equivalent or higher viable cell densities after 24 hours recovery, and up to fi ve-fold higher transient protein expression levels than other media tested.

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•Development Of An Animal-Component Free Electroporation And Recovery Formulation Using EX-CELL™ CHO Cloning Medium

SOURCE: SAFC