14.06.2024
Ready-To-Record cells for manual and automated patch clamp
Cell line maintenance and cell preparation for both conventional and high-throughput patch-clamping remain manual tasks and represent the bottleneck for the kinetic screening of ion channels, in particular voltage-gated ion channels (VGICs).
In their recent book chapter, Emmanuelle Logette and Rajnish Ranjan address this bottleneck by introducing a new approach that significantly enhances the efficiency and scalability of VGIC kinetic screening.
This approach involves the creation of “Ready-To-Record” (RTR) VGIC-expressing cells, which decouples the cell preparation process from patch-clamp experiments.
The study utilizes the Flp-In™-T-Rex™ expression system to create a library of isogenic cell lines (CHO cells) expressing various VGICs. This system ensures site-specific integration and stable expression of ion channel genes, avoiding off-target effects and ensuring high standardization in gene expression.
The RTR CHO cells are then induced with tetracycline to express the ion channels and frozen in specialized vials. The frozen RTR vials can be rapidly thawed and used for both manual and automated patch-clamp recordings within 10 minutes, eliminating the need for continuous cell culture maintenance and streamlining the overall experimental workflow.
This approach offers several significant advantages:
- Efficiency: The ability to freeze and store RTR cells simplifies the logistics of maintaining multiple cell lines, reducing labor and cost.
- Scalability: High-throughput capabilities allow for large-scale kinetic screening, essential for drug discovery and electrophysiological studies.
- Standardization: The use of isogenic cell lines ensures consistency and reliability in experimental results.
Overall, this approach addresses the true limiting factor in ion channel screening, enhancing research efficiency and accelerating drug discovery. It offers a practical solution for electrophysiological studies and drug development processes. The standardized RTR cells ensure a consistent supply, making high-throughput electrophysiological studies more feasible and reliable.
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Find the full book chapter here: Ready-to-Record Cells for Kinetic Screening of VGICs
Learn more about automated patch clamp solutions from Nanion Technologies: https://www.nanion.de/product-categories/automated-patch-clamp/