The proliferation ability of different cancer cells was determined using the xCELLigence Real-Time Cell Analyzer (RTCA)-MP system (Acea Biosciences/ Roche Applied Science) as reported previously 13 (link).
Xcelligence real time cell analyzer mp system
Manufactured by Agilent Technologies
Sourced in United States
The XCELLigence Real-Time Cell Analyzer (RTCA)-MP system is a label-free, real-time cell analysis solution developed by Agilent Technologies. The system continuously monitors changes in electrical impedance, which is correlated to various cellular parameters, such as cell viability, proliferation, and morphology. The RTCA-MP system is designed to provide researchers with a comprehensive and automated platform for monitoring cell-based assays in real-time.
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6 protocols using xcelligence real time cell analyzer mp system
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Real-Time Proliferation Assay for Cancer Cells
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Real-Time Cell Viability Assay of Tumor-PBMC Co-Culture
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Cell viability assays were performed using the xCELLigence Real‐Time Cell Analyzer (RTCA)‐MP system (ACEA Biosciences, Inc., San Diego, CA, USA) as previously described [27 (link)]. Briefly, 1 × 104 cells were seeded into E‐16 plates and placed in the incubator. After 6 h, peripheral blood mononuclear cells (PBMCs) isolated from healthy volunteers were co‐cultured with parental MDA‐MB‐231 cells, shNC‐transfected, or shAURKA‐transfected MDA‐MB‐231 cells at the ratios of 1:4, 1:2, 1:1, 2:1, 4:1, 8:1, and 16:1. Tumor cells cultured without PBMCs were used as control. Cell viability was monitored every minute for 72 h. The cell index was normalized to the starting time of the co‐culture.
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Real-Time Cell Growth Quantification
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Cell growth and proliferation were analysed by growth curves and the Ki-67-positive rate as described previously [54 (link)]. For growth curves, a total of 3 × 103 cells were plated in triplicate in antibiotic-free complete medium in E-Plate 16 (ACEA Biosciences, San Diego, CA, USA) on the xCELLigence Real-Time Cell Analyzer (RTCA)-MP system (ACEA Biosciences) according to the manufacturer’s instructions. The cell index (CI) was read automatically and continuously recorded every hour as CI ± SD. Growth stimulation was determined at the time of maximum cell index (CImax), and a non-parametric t test was used to analyse the significant differences in cell growth. For Ki-67-positive rate analysis, the investigator was blinded to the samples, and ImageJ 1.51j8 (National Institute of Health, USA) was used for analysis.
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Monitoring Cell Proliferation with xCELLigence
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The proliferation ability of KYSE30 and KYSE410 after transfecting with siRNAs and circPUM1 overexpression plasmid was monitored by using the xCELLigence Real-Time Cell Analyzer (RTCA)-MP system (Acea Biosciences/ Roche Applied Science). Transfected cell with siRNAs in six-well culture plate for 24 h, and circPUM1 overexpression plasmid for 48 h, then 50 μL culture medium was added into each well of E-Plate 96 (Roche Applied Science) to obtain equilibrium, and then 3000 cells in 100 μL culture medium were seeded in E-Plate96, which was subsequently locked in RTCA-MP device at 37 °C with 5% CO2. Cell index that reflects cellular proliferation on biocompatible microelectrode coated surfaces was read every 15 min automatically and the recorded curve was shown as cell index±s.e.m.
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Real-Time Cell Proliferation Monitoring
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The proliferation ability of ESCC cells was monitored by xCELLigence Real-Time Cell Analyzer (RTCA)-MP System (Acea Biosciences/Roche Applied Science). Cell index was read automatically every 15 minutes and the recorded curve was shown as cell index AE S.E.M..
6
Real-Time Cell Proliferation Monitoring
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The cell proliferation ability was measured by using the xCELLigence Real-Time Cell Analyzer (RTCA)-MP system (Acea Biosciences, USA), which provided a continuous and quantitative measurement of the cell index to monitor cellular growth status. First, 50 μL of culture medium was added in each well of E-Plate 12 (Roche Applied Science, USA) to detect the baseline. Then cells were suspended in 100 μL medium and seeded in E-Plate 12 with a density of 5×10 4 cells/mL. After 30 min, the E-Plate 12 was inserted into RTCA-MP device and incubated at 37°C with 5% CO 2 . Cell index was measured every 5 min to re ect the cellular proliferation ability.
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