Rtca software pro

Manufactured by Agilent Technologies

The RTCA Software Pro is a software package designed to be used with Agilent's Real-Time Cell Analysis (RTCA) instrumentation. The software provides tools for data acquisition, analysis, and visualization of cellular responses measured using the RTCA system.

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Lab products found in correlation

E plate, by Agilent Technologies (2 mentions) Xcelligence rtca system, by Agilent Technologies (1 mentions) Xcelligence impedance based rtca system, by Agilent Technologies (1 mentions)

2 protocols using rtca software pro

Expansion and Functional Assessment of Engineered CAR T Cells

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For in vitro expansion edited T cells were incubated with EBV-transformed B cells (LCL) in a 1:7 (E:T) ratio and the described protocol was performed accordingly^{33 (link),34 (link)}. CAR frequency and absolute numbers were monitored and measured over indicated time points of expansion.
Sequential stimulation experiment was performed using expanded and rested engineered CAR T cells. Expamers stimulation was done as previously described. Stimulation signal was disrupted by adding 1 mM Biotin. Cells were subsequently washed and transferred into fresh media containing low dose IL-2 (25 IU ml⁻¹). Surface expression of activation markers was monitored by flow cytometry (see below) and cell count was measured (Nucleocounter, Chemotec).
For in vitro killing assay, xCELLigence RTCA System (ACEA Biosciences Inc.) was used. Specific lysis was measured targeting CD19 expressing Human Embryonic Kidney cells (HEK293-CD19⁺). 2 × 10⁴ target cells were seeded into 96 well E-Plate (ACEA Biosciences Inc.) and rested overnight. Engineered CAR T cells were added in a 5:1 (E:T) ratio and incubated for indicated time. Changes in impedance signal were measured in real-time and analyzed using RTCA Software Pro (ACEA Biosciences Inc.). All used cell lines were unmodified and purchased from ATCC.

Fraessle S.P., Tschulik C., Effenberger M., Cletiu V., Gerget M., Schober K., Busch D.H., Germeroth L., Stemberger C, & Poltorak M.P. (2023). Activation-inducible CAR expression enables precise control over engineered CAR T cell function. Communications Biology, 6, 604.

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Real-Time Assessment of NK Cell Cytotoxicity

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The cytotoxicity of NK cells was assessed with a real-time cell analysis (RTCA) assay. Adherent target MCF-7 (3 × 10⁴ cells/well), MDA-MB-231 (3 × 10⁴ cells/well), or SK-BR-3 (4 × 10⁴ cells/well) cells were seeded onto 16-well E-Plates (ACEA Biosciences) in 150 μL of standard RPMI-1640 full medium. For nonadherent K562 cells, 16-well E-Plates were precoated with the Liquid Tumor Killing Assay (anti-CD71) Tethering Kit (ACEA Biosciences) according to the manufacturer's recommendations. K562 cells were seeded onto 16-well E-Plates at a cell density of 1.5 × 10⁴ cells/well. The proliferation of target cells was monitored in the incubator at 37°C (5% CO₂, 95% humidity) for 24 hours with the xCELLigence impedance-based RTCA system (Acea Biosciences). The next day, 100 μL of the medium was aspirated and replaced with the medium containing effector cells (human primary NK cells or NK-92 cell lines) at different effector to target (E:T) ratios. For antibody-dependent cell cytotoxicity (ADCC) assays, trastuzumab (anti-HER2 antibody) was added to appropriate wells at a final concentration of 10 μg/mL. The cells were monitored for the next 20 to 24 hours. Analysis was performed using RTCA Software Pro (ACEA Biosciences). The impedance changes (cell index) were normalized to the end value of the target cells' proliferation and plotted over time as normalized cell index.

Klopotowska M., Bajor M., Graczyk-Jarzynka A., Kraft A., Pilch Z., Zhylko A., Firczuk M., Baranowska I., Lazniewski M., Plewczynski D., Goral A., Soroczynska K., Domagala J., Marhelava K., Slusarczyk A., Retecki K., Ramji K., Krawczyk M., Temples M.N., Sharma B., Lachota M., Netskar H., Malmberg K.J., Zagozdzon R, & Winiarska M. (2021). PRDX-1 Supports the Survival and Antitumor Activity of Primary and CAR-Modified NK Cells under Oxidative Stress. Cancer Immunology Research, 10(2), 228-244.

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