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Breaking HTS Cell-Based Assay Bottlenecks

New Tool Designed to Provide High-Efficiency Transfections with any Molecule in all Cell Types

• James Brady Jr.
• ,
• Karen Donato

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Reproducible Loading

Figure 3. The MaxCyte STX was used to transfect K562 cells with pCMV-eGFP, illustrating its utility for loading suspension cells in large volumes (6x109 cells in 100 mL in ~20 minutes). Click Image To Enlarge +

Figure 3. The MaxCyte STX was used to transfect K562 cells with pCMV-eGFP, illustrating its utility for loading suspension cells in large volumes (6x109 cells in 100 mL in ~20 minutes).

To demonstrate that MaxCyte STX transfections meet HTS requirements for scalability and reproducible loading of agent molecules, MaxCyte transfected six billion K562 cells in about 20 minutes in the MaxCyte STX electroporation buffer containing GFP plasmid DNA.

Figure 3 shows the results of transfecting an aliquot of 40 million cells by small-scale static electroporation and the remainder by large-scale flow electroporation. At 48 hours post-transfection, the large, HTS-scale flow electroporation (red bar) and small-scale, static electroporation (blue bar) yielded almost identical levels of K562 cell viability (>95%) and GFP-positive transfection efficiency (>80%).

Analysis of 28 cell fractions collected at equal time intervals during flow electroporation demonstrated that cell viability and transfection efficiency were highly reproducible and consistent from the beginning through the end of the process.