MRC-5 cells were cultured in DMEM (1×; Thermo Fisher Scientific; 41965039) supplemented with 10% FCS, 100 U/ml penicillin, and 100 µg/ml streptomycin at 37°C and 7.5% CO2. For cytotoxicity assays, MRC-5 cells were infected with varying MOI and subsequently centrifuged for 30 min at 300 g. After resting for ∼1 h, cells were harvested by trypsination for 2 min at 37°C and seeded in E-plates 96 (ACEA Biosciences; 05232368001) with 20,000 cells per well.
E plates 96
Manufactured by Agilent Technologies
Sourced in United States
The E-plates 96 is a specialized lab equipment designed for cell-based assays. It provides a standardized and reproducible platform for monitoring cellular responses in real-time.
Automatically generated - may contain errors
4 protocols using e plates 96
1
MRC-5 Cell Cytotoxicity Assay
2
Cell Growth and Adhesion Monitoring
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Cells were seeded in 200 μl growth medium at a density of 5,000–10,000 cells per well onto E-Plates 96 (ACEA Biosciences). Cell attachment and growth were monitored every 15 min for over 100 hours in the xCELLigence instrument (ACEA Biosciences). Cell growth assays were also performed with the AlamarBlue® cell survival reagent or MTS (Invitrogen, according to manufacturer's instructions), comparable results were observed. For cell adhesion assay, cells were seeded in 200 μl of RPMI medium supplemented with 10% FBS at a density of 5,000–10,000 cells per well onto E-Plates 96. Cell adhesion was monitored every 15 min for 11 hours in the xCELLigence instrument.
3
Quantifying Cell Proliferation Dynamics
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After transfection, cells were seeded into 96-well plates at an intensity of 3,000 cells per well. After the cells adhered to the wall, 10 μl of cell counting kit-8 (CCK-8) detection reagent (Dojindo, Kumamoto, Japan) was added at different time points (24, 48, 72, and 96 h), and the cells were incubated at 37°C for 3 h in the dark. The optical density (OD) at 450 nm was measured using a microplate reader (LabSystems, Vantaa, Finland). Cell proliferation in rescue experiments was done in E-Plates 96 (ACEA Biosciences, San Diego, CA, USA) using a real-time cell analyzer and impedance-based xCELLigence System (RTCA DP Analyzer; Roche Applied Science, Mannheim, Germany). This appliance can monitor cell proliferation dynamically.
4
RPTEC/TERT1 Cell Impedance Monitoring
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If not otherwise indicated, RPTEC/TERT1 cells were seeded on E-plates 96 (ACEA Biosciences Inc., San Diego, CA, USA) at 30% density and placed into the RTCA SP instrument (Roche Diagnostics, Mannheim, Germany). After a growth and maturation period of at least 14 days, cells were treated with compounds or solvent control (0.5% DMSO) or left non-treated. Cellular impedance was measured in intervals of 5 min for a total of 14 days. Treatment was renewed on Mondays, Wednesdays, and Fridays. Cell indices obtained for treated cells were normalized to non-treated cells and are given as percent.
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