neon electroporation transfection system  (Thermo Fisher)

Journal: Scientific Reports

Article Title: Scalable intracellular delivery via microfluidic vortex shedding enhances the function of chimeric antigen receptor T-cells

doi: 10.1038/s41598-025-89070-5

Figure Lengend Snippet: CAR-Ts transfected via hydroporation showed similar viability, proliferation, and EGFR + cells, but improved CAR-T yield, when compared to electroporated or nucleofected cells. ( A ) Overview of workflow for generation of CAR-Ts through RNP and AAV transfection, and how hydroporation is incorporated into the cell therapy workflow with improved cell numbers and viabilities for downstream analysis. ( B ) Picture of the microfluidic hydroporation chip, with an illustration of the microfluidic vortex shedding that cells undergo within the chip (blue window). ( C ) Head-to-head viability comparison of 2 sets of T-cell donors transfected by hydroporation (Hydro; Blue), electroporation (EP; Yellow) or nucleofection (Nuc; Magenta). ( D ) Proliferation of T-cells from day 0 (transfection) to day 9. ( E ) Recovery of T-cells 2 h post transfection. ( F ) Total live T-cells on day 5. ( G ), Percentage of CAR + T-cells, based on EGFR signal. ( H ) Total CAR-T yield on day 5. All data points involve n = 3 technical replicates and, where relevant, p-values from two-tailed heteroscedastic unpaired t-tests.

Article Snippet: Both the Neon Transfection System (electroporation) and the Lonza 4D-Nucleofector (nucleofection) have been adopted as industry standards for intracellular delivery, particularly with hard-to-transfect cell types such as primary human T cells, since there exist unique buffer solutions and pulsing protocols that are optimized for specific cell types , .

Techniques: Transfection, Comparison, Electroporation, Two Tailed Test

Journal: Scientific Reports

Article Title: Scalable intracellular delivery via microfluidic vortex shedding enhances the function of chimeric antigen receptor T-cells

doi: 10.1038/s41598-025-89070-5

Figure Lengend Snippet: Scaled up version of the hydroporation chip, shows similar TRAC-1 KO, viability and recovery when compared to electroporation and nucleofection even when processing 1 billion activated T-cells. ( A ) CAD images of the small volume RUO chip (1x flow cell), standard volume RUO chip (4x Flow cell) and large volume CT chip (40x Flow cell), with an enlarged image showing the post design per flow cell. ( B ) Physical representation of CT chip (40x flow cells, left) to RUO chip (1-4x flow cells, right), bar 5 mm. ( C ) Percentage of CAR + T-cells on day 5 after transfection of 10 6 − 5 × 10 8 total cells. ( D ) TRAC knockout frequency in total live cell population, against cell numbers, based on whether cells were hydroporated (Hydro; blue), electroporated (EP; yellow) or nucleofected (Nuc, Magenta). ( E ) Cell viability 24 h post-transfection. ( F ) Total live cell recovery 2 h post transfection. All data points involve n = 3 biological replicates except 40x FC data.

Article Snippet: Both the Neon Transfection System (electroporation) and the Lonza 4D-Nucleofector (nucleofection) have been adopted as industry standards for intracellular delivery, particularly with hard-to-transfect cell types such as primary human T cells, since there exist unique buffer solutions and pulsing protocols that are optimized for specific cell types , .

Techniques: Electroporation, Transfection, Knock-Out, Cell Recovery