The Use of Fish‐Derived Cell Lines for Investigation of Environmental Contaminants

Dayeh, Vivian R.; Bols, Niels C.; Schirmer, Kristin; Lee, Lucy E. J.

doi:10.1002/0471140856.tx0105s15

Cited by 42 publications

(32 citation statements)

References 27 publications

(46 reference statements)

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“…3, FUs between cultures were analyzed by a one-way ANOVA, and when warranted (p % 0.05), followed by TukeyeKramer post test. For graphical presentation, the results were plotted using SigmaPlot (Jandel Scientific), and EC 50 values were determined using the logistic function option in SigmaPlot for doseeresponse curves [32]. EC 50 values for the different assays were compared by an unpaired t test.…”

Section: Discussionmentioning

confidence: 99%

“…Cytotoxicity was measured as a decline in rezasurin reduction (alamar Blue), a measure of metabolism, and 5-carboxyfluorescein diacetate acetoxymethyl ester (CFDA-AM) conversion to CF, an indicator of plasma membrane integrity [32]. After 30 and 24 h exposures to either the transcriptional inhibitor, actinomycin D (AMD), or translational inhibitor, cycloheximide (CHX), respectively, RTS11 died.…”

Section: Discussionmentioning

confidence: 99%

“…Detailed protocols on the use of these dyes and the interpretation of the results have been described previously [32].…”

Section: Measuring Cell Viabilitymentioning

confidence: 99%

“…Therefore, we have studied the capacity of dsRNA alone or together with inhibitors of macromolecular synthesis to kill the salmonid cell lines, RTS11, RTG-2 and CHSE-214. Cell viability was measured with two fluorescent dyes, alamar BlueÔ and 5-carboxyfluorescein diacetate acetoxymethyl ester (CFDA-AM), which monitor metabolism and cell membrane integrity, respectively [32]. Apoptosis was investigated by looking for characteristic nuclear changes and by determining the effect of the pancaspase inhibitor z-VAD-fmk on cell viability.…”

Section: Introductionmentioning

confidence: 99%

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Preferential induction of apoptosis in the rainbow trout macrophage cell line, RTS11, by actinomycin D, cycloheximide and double stranded RNA

DeWitte‐Orr

Zorzitto

Sutton

et al. 2005

Fish & Shellfish Immunology

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

“…Detailed protocols on the use of these dyes and the interpretation of the results have been described previously [32].…”

Section: Measuring Cell Viabilitymentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Preferential induction of apoptosis in the rainbow trout macrophage cell line, RTS11, by actinomycin D, cycloheximide and double stranded RNA

DeWitte‐Orr

Zorzitto

Sutton

et al. 2005

Fish & Shellfish Immunology

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“…The osmolality of tested solutions were measured three times using a vapor pressure osmometer (Wescor VAPRO 5520). Cells at concentrations around 5×10 5 were seeded into multiwell plates and monitored for morphological changes or for changes in cell number using hemocytometer counts or evaluating viability using Alamar Blue assay as described for fish cell lines (Dayeh et al 2003). To verify that an increase in fluorescence in the Alamar Blue assay is correlated with increased cell density, the assay was completed on two 96-well tissue culture polystyrene plates (Falcon 353072) that contained various cell concentrations of OLGA-PH-J/92.…”

Section: Methodsmentioning

confidence: 99%

Misidentification of OLGA-PH-J/92, believed to be the only crustacean cell line

Lee

Bufalino

Christie

et al. 2011

In Vitro Cell.Dev.Biol.-Animal

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Continuous cell lines from aquatic invertebrate species are few and the development of crustacean cell lines remains an elusive goal. Although a crayfish cell line derived from neural ganglia of Orconectes limosus was reported in 2000, this cell line OLGA-PH-J/92 failed to be authenticated as such. In this report, we describe our attempts to identify the taxonomic identity of the cell line through immunological and molecular techniques. Immunohistochemical screening for the expression of a suite of invertebrate neuropeptides gave negative results, precluding an invertebrate neural origin. PCR amplification and DNA sequencing for the mitochondrial cytochrome c oxydase I, and 18S ribosomal RNA genes that had been widely used to confirm species identity, could not confirm the OLGA-PH-J/92 cells as originating from crayfish. Subsequent attempts to identify the cells provided moderate homology (82%) to Gephyramoeba sp. (AF293897) following PCR amplification of an 18S rDNA fragment after a BLAST search. A literature search provided morphological evidence of the similarity of OLGA-PH-J/92 to the Gephyramoeba distributed by the American Type Culture Collection as ATCC 50654, which also had been misidentified and was renamed Acramoeba dendroida (Smirnov et al., Eur J Protistol 44:35-44, 2008). The morphology of the OLGA-PH-J/92 cells which remains identical to the original report (Neumann et al., In Vivo 14:691-698, 2000) and matched corresponding micrographs that were available from the ATCC before the cell line was dropped from their catalog (ATCC CRL 1494) is very similar to A. dendroida and could thus belong to the Acramoebidae. These results unequivocally indicate that the OLGA-PH-J/92 cell line is not derived from the crayfish O. limosus, and the search for an immortal crustacean cell line continues.

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Delineating cellular interactions between ciliates and fish by co‐culturing Tetrahymena thermophila with fish cells

Pinheiro

Bols

2014

Cell Biology International

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Although several species of Tetrahymena are often described as histophagous and opportunistic pathogens of fish, little is known about ciliate/fish cell interactions, but one approach for studying these is in vitro with cell lines. In this study, T. thermophila, B1975 (wild type) and NP1 (temperature sensitive mutant for phagocytosis) were cultured on monolayers of 3 fish epithelial cell lines, CHSE-214, RTgill-W1, and ZEB2J, and the rabbit kidney epithelial cell line, RK-13. Generally the ciliates flourished, whereas the monolayers died, being completely consumed over several days. The destruction of monolayers required that the ciliates could make contact with the animal cells through swimming, which appeared to dislodge or loosen cells so that they could be phagocytosed. The ciliates internalized into food vacuoles ZEB2J from cell monolayers as well as from cell suspensions. Phagocytosis was essential for monolayer destruction as monolayers remained intact under conditions where phagocytosis was impeded, such as 37°C for NP1 and 4°C for B1975. Monolayers of fish cells supported the proliferation of ciliates. Thus T. thermophila can 'eat' animal cells or be histophagous in vitro, with the potential to be histophagous in vivo.

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The Use of Fish‐Derived Cell Lines for Investigation of Environmental Contaminants

Cited by 42 publications

References 27 publications

Preferential induction of apoptosis in the rainbow trout macrophage cell line, RTS11, by actinomycin D, cycloheximide and double stranded RNA

Preferential induction of apoptosis in the rainbow trout macrophage cell line, RTS11, by actinomycin D, cycloheximide and double stranded RNA

Misidentification of OLGA-PH-J/92, believed to be the only crustacean cell line

Delineating cellular interactions between ciliates and fish by co‐culturing Tetrahymena thermophila with fish cells

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