Preparation of fish primary cell culture

The viruses cause heavy losses in fish farming. Types of cell cultures, including primary cultures are appropriate tools to diagnosis and isolation the viruses. In this study with regard to the appropriate cell lines, especially for fish is not possible, or there are no suitable cells for all fish species was provided, the preparation and standardization of fish primary cell culture were done. Some samples from goldfish (Carassius auratus) internal organs including the kidney, liver, muscles, ovary and external organs including fins, snout connective tissue and gills collected and prepared according to standard procedures. The methods were used include: Enzymatic disaggregated by short and long time trypsinization and without enzyme, by planting minced tissues and ordinary method. The EMEM enriched by FBS was used for growing cells in 25oC incubation. The cells had suitable growth and reached to complete confluency and several subcultures obtained from them. The best result obtained from long time trypsinization of ovaries and muscles. Using the instructions provided in this study can be taken to prepare the primary cell culture for virus isolation and diagnosis of fish viral diseases in the country.