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Long-term primary culture of epithelial cells from rainbow trout (Oncorhynchus mykiss) liver

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Summary

Long-term primary cultures of epithelial cells from rainbow trout (Oncorhynchus mykiss) liver have been established. Nearly homogenous (>97%) populations of hepatocytes were placed into primary culture and remained viable and proliferative for at least 70 d. In addition to hepatocytes, proliferative biliary cells persisted in the cultures for at least 30 d. Finally, a third type of epithelial cell, which we have termed a “spindle cell,” consistently appeared and proliferated to confluence in these cultures. The confluent cultures of spindle cells were successfully subcultured and passaged.

The initial behavior, growth, and optimization of serum and media requirements for these cells is described. All three cell types proliferated as measured by thymidine incorporation, autoradiography, proliferating cellular nuclear antigen analysis, and propidium iodine staining. Further efforts to characterize the cells included western blotting and immunohistochemical staining with antibodies to cytokeratins previously reported in fish liver. From these data, it appears that all three cell populations are epithelial in nature. Furthermore, significant changes in actin organization, often indicative of transformation or pluripotent cells, were observed with increased time in primary culture.

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Authors and Affiliations

  1. Environmental Toxicology Program, Department of Zoology, Oklahoma State University, 74078, Stillwater, Oklahoma

    Gary K. Ostrander & Beverly A. Stark

  2. Department of Biochemistry and Molecular Biology, Oklahoma State University, 74078, Stillwater, Oklahoma

    Gary K. Ostrander & James B. Blair

  3. Cytodiagnostics, Inc., 2925 United Founders Boulevard, 73112, Oklahoma City, Oklahoma

    Garry M. Marley, Wesley D. Bales & Robert W. Veltri

  4. Department of Medicine School of Veterinary Medicine University of California, 95616, Davis, California

    David E. Hinton & Mark Okihiro

  5. Gulf Coast Research Laboratory Ocean Springs, 39566, Mississippi

    Lisa S. Ortego & William E. Hawkins

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  1. Gary K. Ostrander
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Ostrander, G.K., Blair, J.B., Stark, B.A. et al. Long-term primary culture of epithelial cells from rainbow trout (Oncorhynchus mykiss) liver. In Vitro Cell Dev Biol - Animal 31, 367–378 (1995). https://doi.org/10.1007/BF02634286

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