Long-Term Primary Culture of Epithelial Cells from Rainbow Trout (Oncorhynchus Mykiss) Liver

Long-Term Primary Culture of Epithelial Cells from Rainbow Trout (Oncorhynchus Mykiss) Liver

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, proliferat...

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Bibliographic Details
Published in:In vitro cellular & developmental biology. Animal Vol. 31; no. 5; pp. 367 - 378
Main Authors: Ostrander, Gary K., James B. Blair, Beverly A. Stark, Garry M. Marley, Wesley D. Bales, Robert W. Veltri, Hinton, David E., Mark Okihiro, Lisa S. Ortego, Hawkins, William E.
Format: Journal Article
Language:English
Published: Society for In Vitro Biology 01-05-1995
Subjects:
Actins
Antibodies
Cell culture techniques
Cell lines
Cellular Models
Cultured cells
Epithelial cells
Freshwater
Hepatocytes
Keratins
Liver
Liver cells
Marine
Oncorhynchus mykiss
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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 immuno-histochemical 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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ISSN:1071-2690
1543-706X
DOI:10.1007/BF02634286