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Poster: 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.
Poster: Advancing Drug Discovery with the MaxCyte® STX™ Scalable Transient Transfection System: Expression of Intracellular, Membrane-Bound and Secreted Proteins in Physiologically Relevant Cell Lines, Primary Cells and Stem Cells
MaxCyte Platform > Technology > Difficult-to-Transfect Cells
Protein production and most high throughput/ high content screening assays rely on exogenous gene expression such as
reporter genes, expression of fusion proteins, artificially engineered proteins or over expression of a target of
interest. MaxCyte electroporation provides a universal means of cell transfection with the ability to quickly develop
working assays for an extremely broad range of cells including historically difficult to transfect cells. Additionally,
MaxCyte technology can reproducibly transfect up to 1x1010 cells in a single, cost-effective run in less than 30 minutes.
These capabilities allow the MaxCyte system to eliminate reliance on stable cell lines and costly transfection reagents,
and enable rapid, cost-effective performance of cell-based screening and protein production.
The results in the following sections summarize the transient transfection of a variety of difficult to transfect cells using MaxCyte scalable electroporation technology as well as the performance of these cells in a variety of downstream cell-based applications.
The results in the following sections summarize the transient transfection of a variety of difficult to transfect cells using MaxCyte scalable electroporation technology as well as the performance of these cells in a variety of downstream cell-based applications.
Broad Cell-type Compatibility Including Difficult to Transfect Cells
MaxCyte has developed transient transfection protocols for a wide variety of cells including historically difficult to transfect cells. The current diversity and continual expansion of cell-specific Maxcyte electroporation protocols allows for maximum application flexibility as well as future development of more physiological assays and migration of current assays from cell line models to primary cells using a single, universal system. Our list of pre-optimized, cell-specific electroporation protocols is continually being updated. If you don't find your cell type of interest on our list, please contact a MaxCyte scientist for recommendations.
Figure 1 High Efficiency, High Viability Transfection. Ten different cell types were transfected with 200 μg/mL pGFP DNA using the
appropriate pre-loaded protocol. 24 hrs post transfection cells were examined for cell viability (% cells excluding propidium
iodide) and transfection efficiency (% GFP+ cells).
Current MaxCyte STX Protocols
| CHO | Jurkat | Hep G2 | U2OS | RBL | COS-7 |
| Hela | K562 | CV-1 | SH-Sy5y | Neuro2a | LNCaP |
| HEK 293 | NIH 3T3 | THP-1 | COS-1 | NSO | DLD-1 |
| Huh-7 | Renca | Min-6 | A549 | C6 | C2C12 |
| Primary Fibroblasts | Vero | Panc-1 | PC-3 | CaCo-2 | |
| Mesenchymal Stem Cells | PC12 | L5278Y | BHK-21 | RLE |
Nuclear Hormone Receptor Assay using Transfected Jurkat Cells
Nuclear Hormone Receptor Assay using Transfected Jurkat Cells A multi-plasmid reporter gene assay was performed on Jurkat cells transfected using either MaxCyte or other electroporation instrumentation and assay performance compared. Several plasmid ratios were examined as well as a mutant nuclear receptor ligand domain. The nuclear receptor assay sensitivity was found to be 10 times higher using the MaxCyte STX electroporation system compared to another electroporation system.
Figure 2 Electroporation System Comparison for a Nuclear Hormone Receptor Assay. A). Assay Schematic: Cells are co-transfected with a reporter
plasmid (luciferase expressed from a minimal promoter containing multiple GAL4 UAS sequences) and an activator plasmid (constitutively
expressed GAL4 DNA binding domains linked to either a wild-type, WT, or mutant, MT, nuclear receptor ligand binding domain). B).
Cells were transfected using small scale MaxCyte STX electroporation at 2 plasmid ratios (4:1 or 10:1) or using another
electroporation system at a 10:1 plasmid ratio. Cells were seeded in 384-well plates and luciferase activity measured at 5 and
22 hours post electroporation. N = 24 wells.
Antibody Production using CHO Cells
CHO cells are a common cell type used for antibody production, but can be difficult to transfect. The data in Figure 1 summarize results of CHO cell transfection using MaxCyte electroporation. CHO cells were transfected with various concentrations of a mixture of heavy and light chain plasmids and antibody production and cell viability levels examined post transfection. Transiently transfected cells exhibited a high level of viability for an extended period of time as well as produced a high titer of expressed antibody for longer than 10 days. 300 μg/mL plasmid DNA provided the optimum combination of long term cell viability and antibody production.
Figure 3. CHO Antibody Production. CHO cells were transfected with an equimolar mixture of heavy and light chain expression
plasmids on day 0. Cell viability was measured on days 1, 2, and 3 following electroporation. Total IgG concentration in
MaxCyte Outperforms Lipid Reagents
A comparison of CHO cell transfection using MaxCyte electroporation and lipid-based reagents was conducted. CHO cells were transfected with the Kv1.5 ion channel and cells assessed using an automated electrophysiology system. MaxCyte electroporation produced far superior transfection efficiency than lipid-based reagents.
Table 1. Cell Performance in an Automated Ion Channel Assay. CHO K1 cells were transfected with Kv1.5 α-subunit plasmid DNA
using a commercial lipid-based transfection reagent or with MaxCyte EP. Cells were assayed in the single hole and population