Use of trout serum to prepare primary attached monolayer cultures of hepatocytes from rainbow trout (Salmo gairdneri)

Kocal, T. E.; Quinn, B.A.; Smith, Ian R.; Ferguson, H. W.; Hayes, Michael

doi:10.1007/bf02628832

Cited by 32 publications

(19 citation statements)

References 8 publications

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“…A positive influence of lower temperature on longevity of trout cultures was expected. The results support previous in vitro fish studies that showed improved viability at lower temperatures (Klaunig et al, 1985) and enhanced cell attachment (Haschemeyer and Matthews, 1983;Kocal et al, 1988).It has been shown that rainbow trout are sensitive to the cytocidal toxicity of acetaminophen, which requires cytochrome P-450 activation Studies with the medaka from other laboratories have focused on carcinogenesis, with particular emphasis on the histopathological identification of neoplastic lesions in the tissues of animals exposed to carcinogens. Because a long latent period often occurs between initial contact with deleterious chemicals and subsequent expression of the pathology, we are investigating early biologically relevant responses that can be used as genotoxicity markers of exposure and effect.…”

supporting

confidence: 87%

“…The initial strategy was to culture cells in a serum-free medium to allow the study of cellular behavior under chemically defined and controlled conditions. However, since serum was used as a medium supplement in some trout studies (Lipsky et al, 1986;Kocal et al, 1988), the investigators tested the effect of 10% fetal bovine serum in their system. Based on their preliminary studies in which fetal bovine serum showed no effect on cell attachment, 30 spreading, and viability over time, these investigators have eliminated it from their medium components.…”

mentioning

confidence: 99%

“…Every 24 hours thereafter, the medium was removed, and the cultures were treated again. Four culture dishes per treatment level were used.Vehicle control cultures were exposed to 0.4% DMSO (v/v).As an indicator of retained function and hepatic differentiation, lactate dehydrogenase (LDH) leakage due to cytochrome P-450 activation by acetaminophen Kocal et al, 1988) was assayed using a Sigma LDH assay kit at 25°C (Wroblewski and LaDue, 1955).Trypan blue exclusion was also used as an indicator of viability, which remained at 85 ± 5% throughout the experiment. …”

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confidence: 99%

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Compendium of the FY1990 and FY1992 Research Reviews for the Research Methods Branch

Gardner¹

1994

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Public reporting burden for this collection of information is estimated to average I hour per response, including the time for reviewing Instructions, searching existing data sources. gathering and maintaining the data needed, and completing and reviewing the collection of information Send comments regarding this burden estimate or any other aspect of this collection of information, including suggestions for reducing this burden, to Washington Headquarters Services, Directorate Approved for public release; distribution unlimited ABSTRACT (Maximum 200 words)Since 1983, the U.S. Army Biomedical Research and Development Laboratory (USABRDL) has conducted a research program focused on the development of new methods for assessing the potential hazards resulting from exposure to chemicals in the workplace or the environment. This research has been conducted both in-house and extramurally and has involved scientists from other Federal agencies, academic institutions, and the private sector. The need to enhance the integration of the research became increasingly evident as the program grew in the late 1980's. This growth came principally as a result of the U.S. Army Corps of Engineers sponsorship of research to develop environmental applications of new, in vivo hazard assessment models for use in the burgeoning Department of Defense environmental remediation program. One of the many uses of these models is to answer the question, "How clean is clean?" and to assist in the establishment of more realistic clean-up goals for hazardous waste sites.An annual research review meeting was undertaken to facilitate the integration and technology transfer of this research program. This publication is the second 14. SUBJECT TERMS Fish, Medaka, Irout, #13.(Continuation) compendium of these meetings.The papers contained in this document are meant to provide the program management staff with an additional means of reviewing the status and progress of extramural and selected in-house studies in the area of new toxicity assessment models. The papers are, therefore, the principal investigators' (PIs') best efforts to characterize the research at the time of the meeting.Some papers cover the results of several years of research and others outline only the planned research of a new project. The data presented and conclusions of the authors represent the best professional judgment of the respective PI at the time of the workshop but are not equivalent to a peer-reviewed report of completed research. NOTICE DisclaimersThe views, opinions, and/or findings contained in this report are those of the author(s) and should not be construed as official Department of the Army position, policy, or decision, unless so designated by official documentation.Citations of commercial organizations or trade names in this paper do not constitute an official Department of the Army endorsement or approval of the products or services of these organizations.Research was conducted in compliance with the Animal Welfare Act, and other Federal statutes and regulati...

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confidence: 87%

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confidence: 99%

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confidence: 99%

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Compendium of the FY1990 and FY1992 Research Reviews for the Research Methods Branch

Gardner¹

1994

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“…In oviparous animals, the synthesis of VTG by hepatocytes represent a good system with which to study the mechanism of action of steroid hormones [31][32][33][34]. Most of these studies have used hepatocytes of either amphibians (especially Xenopus laevis) or birds (especially chickens), though some have been based on hepatocytes from fish [24,[35][36][37]. Various techniques have been used to culture fish hepatocytes [35,37].…”

Section: Resultsmentioning

confidence: 99%

“…Most of these studies have used hepatocytes of either amphibians (especially Xenopus laevis) or birds (especially chickens), though some have been based on hepatocytes from fish [24,[35][36][37]. Various techniques have been used to culture fish hepatocytes [35,37]. We chose to use the system first developed by Flouriot and Valotaire [38] and modified by Foucher et al [39], in which gentle shaking is used to encourage the hepatocyte to form small aggregates, as a first step towards tissue organization [40].…”

Section: Resultsmentioning

confidence: 99%

Vitellogenin synthesis in cultured hepatocytes; an in vitro test for the estrogenic potency of chemicals

Pelissero

Flouriot

Foucher

et al. 1993

The Journal of Steroid Biochemistry and Molecular Biology

266

108

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Smnmary--We describe here an/n vitro technique to assess the estrogenic activity of chemicals. This technique is based on rainbow trout hepatocytes incubated in a basic medium free of any additional growth factors or estrogenic chemicals and uses the production of vitellogenin (VTG) as a marker for the estrogenic potency of the compounds tested. The system allows at least some of the metabolic transformations which are undertaken by the liver cells/n vivo and could therefore be used for xenobiotic compounds which exhibit estrogenic activities after liver metabolic transformation. A dose-response curve was always consistently obtained using estradiol-17fl (E2), with a mid point at around 100 nME 2 and a maximum response at around 1000nM. Established estrogens such as 17al ethynylestradiol (EE2) or diethylstilboestrol (DES) were also tested. EE 2 appeared to be equipotent with E2 and DES slightly less potent. E2 conjugates were, perhaps surprisingly, also very potent. Estradiol-3-sulfate was equipotent with E2 and estradiol-17fl-glucuronide approx. 10% as potent. Other steroids such as androgens and progesterone, though active in the bioassay, were 3 orders of magnitude less potent than E 2. Of the various steroids tested, only cortisol, at concentrations up to 50 #M, was completely inactive. Six different phytoestrogens were tested in the assay. All were weakly estrogenic, possessing approximately one thousanth the potency of E 2 (they were as potent as the androgens and progesterone). All six phytoestrogens, as well as the androgens and progesterone, were tested in the presence of tamoxifen. In all cases tamoxifen reduced the production of VTG significantly, demonstrating that the estrogenic action of all of these compounds was most likely mediated by the E 2 receptor. The potencies determined here may not reflect the situation/n vivo but can provide complementary results about the activity of chemicals which need an hepatic metabolization to be estrogenic. Hepatocyte cultures would profitably be developed in other species to sustain these results.

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Recent applications of fish cell culture to biomedical research

1988

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Tissues of the fishes are as amenable to the techniques of modern cell culture as mammalian tissues and organs, and yet this vast resource, comprising thousands of vertebrate species, remains largely unexplored. The model systems that have been developed demonstrate the utility of fish cells as sources of special adaptations and exaggerated physiological systems. In this review, we briefly describe several of the successful models along with recent developments in fish cell culture with the hope of stimulating increased interest in the lower vertebrates as useful complements to mammalian cell culture in biomedical research. The topics covered include epithelial ion transport, endocrinological studies, the cellular stress (heat shock) response, thermotolerance, cancer biology, and environmental toxicology.

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Use of trout serum to prepare primary attached monolayer cultures of hepatocytes from rainbow trout (Salmo gairdneri)

Cited by 32 publications

References 8 publications

Compendium of the FY1990 and FY1992 Research Reviews for the Research Methods Branch

Compendium of the FY1990 and FY1992 Research Reviews for the Research Methods Branch

Vitellogenin synthesis in cultured hepatocytes; an in vitro test for the estrogenic potency of chemicals

Recent applications of fish cell culture to biomedical research

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