About
Standard passaging of adherent HEK-293T cells typically relies on centrifugation to pellet cells and wash away residual dissociation reagent. While centrifugation can be automated, removing it from the workflow reduces process complexity, cuts hands-on time, and increases throughput. This application note shows how centrifugation-free passaging can maintain healthy HEK-293T cultures when residual dissociation reagent is kept below a defined threshold, and demonstrates the approach running unattended on the Cellmatic system across multiple passages. Inside, you'll find the tolerance data, the automated protocol, and the growth curves.
The Challenge: Why Centrifugation Holds Back Cell-Culture Automation
HEK-293T cells are a workhorse for transient transfection, recombinant protein production, and viral-vector generation, but they adhere loosely and are easily disturbed by repeated handling. Conventional passaging relies on aspiration, washing, and centrifugation, a sequence that is labor-intensive and time-consuming. Eliminating the centrifugation step reduces process complexity and increases throughput, but only if the residual dissociation reagent carried into the daughter culture stays low enough to protect cell attachment and growth.
What's Inside This Application Note
- The residual dissociation-reagent (1X trypsināEDTA) tolerance threshold for HEK-293T cells, established across a 0ā25% (v/v) carryover range
- Why a single overnight media exchange rescued culture integrity even at the highest carryover tested
- The centrifugation-free automated protocol used on the Cellmatic, and the residual level it achieves
- Passage-by-passage growth and morphology data over long-term maintenance
- Practical strategies to keep carryover low in higher-split-ratio workflows
Automated centrifugation-free maintenance of HEK-293T culture
FAQs
It's a passaging method where cells are subcultured directly using a diluted enzyme/media mixture, without a centrifugation step to pellet cells and wash away the dissociation enzyme before reseeding. This significantly increases throughput and reduces complexity in automated workflows, reduces cell handling time, and minimizes mechanical shear stress on sensitive cells.