Primary Cell Data
Opening New Horizons - Efficient 96-well Transfection of Primary Cells
Transfection methods such as lipofection or electroporation yield satisfactory results for standard cell lines only, whereas many cell lines and especially primary cells are difficult or even impossible to transfect with conventional methods. Using the Nucleofector™ 96-well Shuttle™ System, primary cells can be reliably transfected with high efficiency and high viability in a high throughput format. Thereby, applications such as target validation can now for the first time be performed in primary cells, hence drastically increasing result reliability.
- High convenience - Optimized protocols for each cell type with details on cell isolation and culture
- Flexible cell numbers - Between 104 and 106 cells in 20 µl transfection volume
- Access to difficult-to-transfect cell types - Neurons, T cells and other difficult-to-transfect cells types can be transfected
- Short time-to-results - Expression within hours allows transfection and analysis in one day
Example for the transfection of primary cells using the 96-well Shuttle™. Cells were transfected with 400 ng pmaxGFP™ Vector and analyzed for maxGFP™ Expression on a Becton Dickinson FACSCalibur with HTS option, 24h post Nucleofection™. Cell viabilities range between 30% and 92% depending on the cell type.
Cell Line Data
The ideal tool for transfection of cell lines for screening applications
The Nucleofector™ Technology can be used for transfection of virtually any type of cell line such as commonly-used cell lines or difficult-to-transfect cell lines such as suspension cells. High transfection efficiencies combined with high cell viability and the parallel 96-well approach make it the ideal tool for cDNA or RNAi library screening. The can be used for transfection of virtually any type of cell line such as commonly-used cell lines or difficult-to-transfect cell lines such as suspension cells. High transfection efficiencies combined with high cell viability and the parallel 96-well approach make it the ideal tool for cDNA or RNAi library screening.
- High efficiencies - Up to 99% siRNA transfer efficiency and up to 95% DNA transfection efficiency
- Fast and easy optimization - Optimization of transfection conditions for virtually any cell line within one plate
- High convenience - Optimized protocols available for cell lines with details on cell isolation and culture
- Access to difficult-to-transfect cell lines - Transfection of suspension cell lines
- Ideal for high throughput applications e.g. RNAi library screens
Highly efficient transfection of various cell lines using the 96-well Shuttle™ System. Cell lines were transfected with 400 ng pmaxGFP™ Vector and analyzed for maxGFP™ Protein Expression on a Becton Dickinson FACSCalibur with HTS option, 24h post Nucleofection™. Cell viabilities range between 50% and 97% depending on the cell line.
New potentials in target validation: RNAi library screening using difficult-to-transfect cell lines
A general trend towards the use of primary cells and challenging cell lines demands a highly efficient and reliable transfection technology. This is especially true for successful RNAi library screens (such as in target validation studies) where efficient transfection of cells with siRNA duplexes or shRNA vectors is essential. The Nucleofector™ Technology provides unique access to primary cell and cell line based siRNA screens delivering data of outstanding quality.
siRNA-mediated depletion of vimentin-mRNA in Jurkat cells. Jurkat cells (2x105) were transfected with an siRNA duplex directed against endogenous vimentin. 24h post transfection vimentin mRNA levels were analyzed by RealTime PCR. Relative expressions compared to untreated control sample (well 31, set to 100%) are shown. (External data kindly provided by C. Merz, Schering AG, Berlin).