High Throughput Transfection of Stem Cells, Primary Cells and Difficult-to-Transfect Cell Lines: Jurkat, CHO, Human Skeletal Muscle Cells & Primary Neuronal Cell Transfection using a Scalable, Electroporation-Based Technology
Most high throughput, high content screening and profiling assays rely on exogenous gene expression of reporter genes, fusion proteins and artificially engineered proteins or overexpression of a target of interest. While stable cell lines are a convenient tool for producing the large number of cells required to perform a single screen, they are time-, labor- and cost-intensive to create. Transient transfection offers the ability to quickly develop working assays, however, many of these technologies have limitations on compatible cell types. Additionally, they can require multiple small-scale transfections or use costly transfection reagents to produce a large number of cells. MaxCyte® scalable electroporation offers a cost-effective, reproducible transfection of up to billions of cells in less than 30 minutes with broad cell type compatibility. In this poster we present the transient transfection of a variety of difficult-to-transfect cells and their use in downstream assays. Specifically, we demonstrate how large-scale electroporation of Jurkat, CHO, human skeletal muscle cells and primary neuronal cells produces large numbers of quality transfected cells and can ease the burden of stable cell line engineering.
MaxCyte STx® Scalable Transfection System. MaxCyte offers a proprietary, scalable electroporation technology to transfect a variety of cell types, including primary cells, with DNA, RNA, siRNA, proteins or other biomolecules of interest. MaxCyte instruments feature optimized electroporation (EP) protocols for a wide range of cell types, simplifying assay development while maximizing performance and reproducibility. For a wide range of cell types, transfection efficiencies exceed 85% and cell viability is better than 90%. The MaxCyte STx can perform small-scale transfections of 5x105 cells in seconds for use in basic research and assay development, or in less than 30 minutes for bulk transfections of up to 1x1010 cells in full-scale screening and profiling experiments.
- High efficiency
- Broad cell type compatibility
Current MaxCyte STx® Protocols
- NIH 3T3
- Hep G2
- L5278Y OS
- HEK 293
- Primary Fibroblasts
- Mesenchymal Stem Cells
Nuclear Hormone Receptor Assay
|MaxCyte Electroporation||EC 50||S/B|
|Small-Scale||2.39E - 06||635|
|Large-Scale||2.60 E - 06||604|
High Throughput Ion Channel Assay
|Lipid Transfection (20 µg DNA + 60 µl lipid reagent)||77%||191±46 MΩ||4%||1.1±1.0 nA|
|MaxCyte Electroporation (150 µg/ml cDNA, 48 hr post-electroporation)||Single Hole||82%||248±87 MΩ||93%||2.8±1.4 nA|
|Population Patch Clamp||100%||72±31 MΩ||98%||1.3±0.30 nA|
Primary Cell Transfection
Human Skeletal Muscle Cell Transfection
Neuronal Cell Transfection
- MaxCyte electroporation can be used to transiently (co)transfect a variety of primary cells, stem cells and historically difficult-to-transfect cell lines with high transfection efficiencies and cell viability.
- The MaxCyte STx transfection system is fully scalable, allowing researchers to rapidly transfect from 5x105 to 1x1010 cells using the same protocol.
- MaxCyte electroporation can be used to express a variety of functional proteins including secreted proteins, membrane proteins and nuclear proteins.
- Cells transfected using the MaxCyte STx produce quality results when used in a variety of cellular assays and in antibody protein production.