Establishment and comparative analyses of different culture conditions of primary hepatocytes from Nile tilapia (Oreochromis niloticus) as a model to study stress induction in vitro |
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Authors: | Joseph O Jung'a Eric S Mitema Herwig O Gutzeit |
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Institution: | 1. Institute of Zoology, Faculty of Mathematics and Natural Sciences, Technische Universit?t (TU), 01062, Dresden, Germany 2. Institute of Primate Research, National Museums of Kenya, P.O. Box 24481, 00502, Karen, Kenya 3. Faculty of Veterinary Medicine, University of Nairobi, P.O. Box 30197, 00100, Nairobi, Kenya
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Abstract: | Summary We established an in vitro hepatocyte primary culture system from Oreochromis niloticus, a tropical fish species of great economical importance, and evaluated its ability to express albumin, a liver-specific protein,
consistently for a period of 3 wk. Serum requirements for fish hepatocyte cultures were assessed. A one-step in situ perfusion
of tilapia liver retrogradely followed by collagenase liver dissociation and subsequent washing produced nearly 90% homogenous
viable hepatocytes, as shown by trypan blue exclusion test. Mixed primary monolayer and aggregate hepatocyte cultures achieved
by 10% fetal calf serum medium supplements expressed consistent levels of albumin. The results of light and electron microscopy
showed that the hepatocytes did not significantly proliferate (P<0.05) but remained viable for at least 3 wk. The results of this study show that in vitro cultures of mixed primary hepatocyte
monolayers and aggregates established from Nile tilapia may be useful models for studying transient cellular stress induction. |
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Keywords: | primary hepatocytes bioassay model system transient cellular stress expression Nile tilapia HSP70 |
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