Xcelligence rtca s16

Proliferation of RAW 264.7 cells, as obtained from American Type Culture Collection (ATCC TIB-71) was monitored with Real Time Cellular Analysis system (xCELLigence RTCA S16, ACEA Biosciences, San Diego, USA) in Dulbecco’s modified Eagle’s medium (DMEM, Caisson, Smithfield, UT, USA) containing 10% fetal bovine serum (FBS, Genedirex, Las Vegas, NV, USA) with 1% penicillin/streptomycin in a humidified atmosphere with 5% CO₂ in a 37 °C incubator. The toxicity assays of both methotrexate and diclofenac sodium were performed with RTCA system at a series of geometric concentrations (1 mM, 100 μM, 10 μM, 1 μM, 100 nM, 10 nM, 1 nM, 0 fM). The non-toxic dosages of methotrexate and diclofenac sodium were selected to investigate their anti-inflammation effect. 3500 cells in every well were cultured in the E-plates of the RTCA system and stimulated with lipopolysaccharide (LPS, 1 μg/mL, Sigma, Darmstadt, Germany. Cat. L2630) at 12th h for 4 h. After stimulating with LPS, the medium of Model group was continued to be under the stimulation of LPS. The anti-inflammation groups were treated with methotrexate in different concentrations or diclofenac sodium respectively. Control was cultured with 10% DMEM+PBS after the four hours’ stimulation with LPS.

The inhibitory effects of the genome-edited NK cells on GBM cells were investigated using the xCELLigence RTCA S16 and DP instruments (ACEA Biosciences, San Diego, CA, USA). The procedure has been described previously [55 (link)]. Briefly, complete medium (100 μL) was added to each well on E-plate 16 (ACEA Biosciences), and background impedance was measured at 37 °C in a humidified atmosphere containing 5% CO₂. The GBM cells, i.e., 2 × 10⁴/well T98G or LN-18cells, were seeded in each well as the target cells, and impedance measurement was recorded every 5 min for 72 h. After 24 h, the genome-edited NK cells were added to each well as the effector cells in the defined E:T cell ratios. The data were analyzed using RTCA, version1.2 (ACEA Biosciences).

Real-time cell analysis (RTCA) was conducted using the xCELLigence RTCA S16 (ACEA, San Diego, USA). Briefly, 50 μL of the medium was added to 16-well E-Plates to obtain background readings before adding 100 μL of the cell suspension (4 × 10⁴ cells/well). After incubation at room temperature for 30 min, the E-Plates were placed on the reader in an incubator, and the cell index recording continued. After 24 h, the cells were treated with sanguinarine at different concentrations. Cells were monitored every 15 min for 48 h after treatment.

Real-time cell analysis (RTCA) is a technology based on the principle of the microelectronic biosensor chip, which can realize the real-time analysis of cells without markers in the process of experiments [21 (link)]. The RTCA instrument (xCELLigence RTCA S16; ACEA Biosciences, SD, CA, USA) was used to analyze the proliferation and adhesion properties of primary cultured trophoblasts of mice. Briefly, 50 μL of culture medium was added to the well of an E16 xCELLigence microtiter plate, which was then inserted into the RTCA device. After 1 min, the background impedance was measured for each well. Subsequently, trophoblasts of mice (1 × 10⁴ cells in 50 μL culture medium) were added to each well and cultured for 2 h and then treated with different concentrations of FST for another 72 h in the proliferation assay. In the adhesion assay, trophoblasts of mice and FST were added to each well together and cultured for 5 h. Cells were all monitored in 15 min intervals. The cell index (CI) reflected the biological status of monitored cells, including the cell number, viability, morphology and adhesion.

Real-time cell analysis (RTCA) is a technology based on the principle of microelectronic biosensor chip, which can realize the real-time analysis of cells without markers in the process of experiment. The RTCA instrument (xCELLigence RTCA S16; ACEA Biosciences, California, United States) was used to analyze the proliferation and adhesion properties of L929 cells. Initially, 50 μl of cell-free culture medium was added to the well of an E16 xCELLigence microtiter plate, which was then inserted into the RTCA device. After 1 min, the background impedance was measured for each well. Subsequently, L929 cells (2 × 10⁴ cells in 50 μl culture medium) were added to each well and cultured for 3 h, and then treated with different concentrations of activin A for another 67 h in the proliferation assay. In the adhesion assay, cells and activin A were added to each well together and cultured for 4 h. Cells were monitored in 5 or 15 min intervals. Each experiment was performed in duplicate and repeated three times. The impedance of the cell sensor was described and measured as the cell index (CI), which reflected the cell activity.