Xcelligence rtca sp system

HeLa cells were exposed to heat shock at 45°C for 0, 7.5 and 15 min and recovered at 37°C for 3 h in fresh media. For real-time cell analyzer analysis, 10,000 cells were plated in 96 well E-plate with 200 μL of media and cell growth was monitored by measuring electrical impedance every 15 min under xCELLigence RTCA SP system (Roche Applied Science). WST-1 cell proliferation assay was performed using WST-1 (Roche). HEK293T cells were transfected with Flag or Flag-hArc vector and treated with heat shock at 45°C for 0, 7.5 and 15 min, then cells were dispensed in 96-well microtiter plates and incubated at 37°C with fresh media for 1 day, and cell survival was measured using WST-1 solution. The cleavage of WST-1 to formazan by metabolically active cells was quantified by scanning the plates in a microtiter plate reader at 440 and 620 nm (reference wavelength). The test medium was used as the background control. Three independent sets of experiments that were performed in triplicate were evaluated. The viability of the treated cells was normalized to the untreated control cells.

The xCELLigence RTCA SP system (Roche Applied Science; Mannheim, Germany) was used for real-time and time-dependent analysis of the cellular response of thyroid cancer cells following incubation with 1 nM to 10 µM panobinostat, 1 nM to 10 µM SAHA, and 1 nM to 10 µM Trichostatin A. To perform this analysis, 3000 cells were seeded in 150 μL complete growth medium per well in a 96-well E-plate (OLS; Bremen, Germany) and were incubated with the previously mentioned compounds. Cell Index, which indicates attachment and adherence of cells to the plate’s electrode, was continuously measured for the following 120 h using the cell impedance detection system. Data analysis was performed using the RTCA Software v1.2.1 for the calculation of the temporal dynamics of cellular attachment (i.e., viability).

Proliferation curves from ORL cell lines and NOK cells were generated using the xCELLigence RTCA SP system (Roche Applied Science, Upper Bavaria, Germany). Cell proliferation was monitored every 15 minutes for the initial 2 hours and then every 30 minutes for the subsequent 120⁻166 hours. The start and end time during the log-growth phase of each cell lines were selected and doubling time was calculated by RTCA software (Roche Applied Science, Upper Bavaria, Germany). Doubling time was calculated from 2–3 independent experiments, with at least triplicate wells per experiment.

NC cells were seeded in E-96-well plates and infected 24 h later with T-VEC. At 1 hpi distinct SMIs were added. Real-time dynamic cell proliferation was measured every 30 min during the full observation time of 96 h by xCELLigence RTCA SP System (Roche Applied Science, Penzberg, Germany). The cell impedance, i.e., the electrical resistance to alternating current, was expressed in an unspecified unit as the cell index. Cell index values were calculated using the RTCA software (1.2.1; Roche Applied Science, Penzberg, Germany).

H727 cells were seeded in 96-well plates (E-Plate 96, Roche Applied Science, Mannheim, Germany). The xCELLigence RTCA SP system (Roche Applied Science) was employed to observe impedance of the cell layer in 30 min intervals over 120 h. 24 h after seeding, cells were infected with GLV-1 h68 using MOIs 0.1 and 0.25 or treated with 0.1% (v/v) Triton for lysis control. The measured impedance was used to calculate Cell Index values with the RTCA Software (1.0.0.0805).

A673 cells (5 × 10³ cells/well) were seeded in 96-well plates (E-Plate 96, Roche Applied Science, Mannheim, Germany). Real-time dynamic cell proliferation was monitored in 30 min intervals during a 130 h observation period using the xCELLigence RTCA SP system (Roche Applied Science). Cell index values were calculated using the RTCA Software (1.0.0.0805). 21 h after seeding, cells were infected with MeV-GFP at MOI 0.5 or mock-infected. At 51 hpi PBMC, PBMC stimulated with IL-2 or NK cells were added at the indicated effector to target (E:T) ratios [24 (link), 25 (link)]. HT1080 cells (1 × 10³ cells/well) were infected at 24 h after seeding with MeV-GFP at MOI 5 or mock-infected. At 23 hpi, NK cells were added at E:T ratios ranging from 1:1 to 5:1. Cell proliferation was monitored in 60 min intervals during a 96 h observation period.

The xCELLigence RTCA SP system (Roche Applied Science) was used, for real-time analysis of the cellular response of HepG2 and Hep3B cells following incubation with 100 nM panobinostat, 5–10 μM 4-HidroxyTamoxifen, 100 nM 1 μM 3-methyladenine and 10–100 pM Bafilomycin as previously described [19 (link)–21 (link)]. Cell index, indicating attachment and adherence of cells to the plate's electrode, was measured continuously for the following 80 hours. Data analysis was performed using the RTCA Software v1.2.1.