Monitoring viral-induced cell death using electric cell-substrate impedance sensing.

Using an electrical measurement known as electric cell-substrate impedance sensing (ECIS), we have recorded the dynamics of viral infections in cell culture. With this technique, cells are cultured on small gold electrodes where the measured impedance mirrors changes in attachment and morphology of cultured cells. As the cells attach and spread on the electrode, the measured impedance increases until the electrode is completely covered. Viral infection inducing cytopathic effect results in dramatic impedance changes, which are mainly due to cell death. In the current study, two different fish cell lines have been used: chinook salmonid embryonic (CHSE-214) cells infected with infectious pancreatic necrosis virus (IPNV) and epithelioma papulosum cyprini (EPC) carp cells infected with infectious hematopoeitic necrosis virus (IHNV). The impedance changes caused by cell response to virus are easily measured and converted to resistance and capacitance. An approximate linear correlation between log of viral titer and time of cell death was determined.

[1]  Ari Helenius,et al.  How Viruses Enter Animal Cells , 2004, Science.

[2]  M. Soler,et al.  Attachment and entry of infectious pancreatic necrosis virus (IPNV) into CHSE-214 cells , 2005, Archives of Virology.

[3]  B. Nicholson,et al.  Studies on a host range variant from different isolates of infectious pancreatic necrosis virus (IPNV) , 1979 .

[4]  I. Giaever,et al.  Electric measurements can be used to monitor the attachment and spreading of cells in tissue culture. , 1991, BioTechniques.

[5]  I. Giaever,et al.  Combining optical and electrical impedance techniques for quantitative measurement of confluence in MDCK-I cell cultures. , 2004, BioTechniques.

[6]  T. Lin,et al.  Apoptosis precedes necrosis of fish cell line with infectious pancreatic necrosis virus infection. , 1998, Virology.

[7]  Ivar Giaever,et al.  A morphological biosensor for mammalian cells , 1993, Nature.

[8]  E. Everitt,et al.  Temporal and subcellular localization of infectious pancreatic necrosis virus structural proteins , 2000, Archives of Virology.

[9]  K. Leung.  Current trends in the study of bacterial and viral fish and shrimp diseases , 2004 .

[10]  Eugenia Wang,et al.  Use of electric cell-substrate impedance sensing as a tool for quantifying cytopathic effect in influenza A virus infected MDCK cells in real-time. , 2005, Journal of virological methods.

[11]  S. Lapatra,et al.  DNA vaccines for aquacultured fish. , 2005, Revue scientifique et technique.

[12]  D. Amend.  Control of infectious Hematopoietic Necrosis virus disease by elevating the water temperature , 1970 .

[13]  C.R. Keese,et al.  A biosensor that monitors cell morphology with electrical fields , 1994, IEEE Engineering in Medicine and Biology Magazine.

[14]  C. Lo,et al.  Monitoring motion of confluent cells in tissue culture. , 1993, Experimental cell research.

[15]  Joachim Wegener,et al.  Bioelectrical impedance assay to monitor changes in cell shape during apoptosis. , 2004, Biosensors & bioelectronics.

[16]  J. C. Espinoza,et al.  Infectious pancreatic necrosis virus (IPNV) does not requireacid compartments for entry into cells , 1997, Archives of Virology.

[17]  I. Giaever,et al.  Monitoring fibroblast behavior in tissue culture with an applied electric field. , 1984, Proceedings of the National Academy of Sciences of the United States of America.

[18]  J. Wegener,et al.  Electric cell-substrate impedance sensing (ECIS) as a noninvasive means to monitor the kinetics of cell spreading to artificial surfaces. , 2000, Experimental cell research.

[19]  K. Wolf.  Fish Viruses and Fish Viral Diseases , 2019 .

[20]  H. Miller,et al.  Fish cell lines: comparisons of CHSE-214, FHM, and RTG-2 in assaying IHN and IPN viruses. , 1978 .

[21]  J. Fryer,et al.  THE IN VITRO CULTIVATION OF TISSUE AND CELLS OF PACIFIC SALMON AND STEELHEAD TROUT * , 1965, Annals of the New York Academy of Sciences.

[22]  Petr Skládal,et al.  Piezoelectric biosensors for real-time monitoring of hybridization and detection of hepatitis C virus. , 2004, Journal of virological methods.

[23]  T. Yamamoto,et al.  Purification of infectious pancreatic necrosis (IPN) virus and comparison of polypeptide composition of different isolates. , 1978, Canadian journal of microbiology.

[24]  J. Fryer,et al.  Properties of the Sockeye Salmon Virus (Oregon Strain)∗ , 1969, Proceedings of the Society for Experimental Biology and Medicine. Society for Experimental Biology and Medicine.

[25]  J-R Hong,et al.  Induction of apoptotic death in cells via Bad gene expression by infectious pancreatic necrosis virus infection , 2002, Cell Death and Differentiation.

[26]  E. Hm,et al.  Comparison of representative strains of infectious hematopoietic necrosis virus by serological neutralization and cross-protection assays. , 1991 .

[27]  Ivar Giaever,et al.  Use of Electric Fields to Monitor the Dynamical Aspect of Cell Behavior in Tissue Culture , 1986, IEEE Transactions on Biomedical Engineering.