Cellular Impedance-Based Survival Kinetics Assay to Assess Virus Survival

Abstract

Source: Labadie, T. et al., Monitoring Influenza Virus Survival Outside the Host Using Real-Time Cell Analysis. J. Vis. Exp. (2021).

This video describes a cellular impedance-based technique to study the survival kinetics of viruses that induce cytopathic effects. The method measures the electrical impedance of virus-infected adherent cells pre-treated with saline.

Protocol

1. Preparation of reagents and starting materials

1. Preparation of MDCK cells and sterile cell culture medium
   1. Cultivate Madin-Darby Canine Kidney (MDCK) cells in modified Eagle's medium (MEM) supplemented with 10% heat-inactivated fetal calf serum (FCS) and antibiotics (100 units/mL penicillin, 100 mg/mL streptomycin).
   2. After thawing, passage MDCK cells at least 2x before infecting them to ensure complete recovery (but less than 30 passages to avoid any drift in cell phenotypes).
   3. Seed 75 cm²  tissue culture flask(s) containing 30 mL of sterile 1x MEM with 7.5 x 10⁶  MDCK cells and incubate at 3....

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
0.25% Trypsin	ThermoFisher	25200056
75 cm2 tissue culture flask	Falcon	430641U
E-Plate 16 (6 plates)	ACEA Biosciences, Inc	5469830001	E-plates are available in different packaging
FCS	Life technologies (gibco)	10270-106
MEM 1X	Life technologies (gibco)	31095029
PBS 1X	Life technologies (gibco)	14040091
Penicillin-Streptomycin	Life technologies (gibco)	11548876
TPCK-Trypsin	Worthington	LS003740
xCELLigence Real-Time Cell Analysis Instrument S16	ACEA Biosciences, Inc	380601310	The xCELLigence RTCA S16 instruments are available in different formats (16-well, 96-well, single or multi-plate