Primary Fish Cell Culture Phase 1: primary cell culture of the interior pituitary gland of the Acipenser gueldenstaedtii
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AbstractFish cell lines have been used to support many areas of research, beginning with fish viruses and extending into immunology, genetic studies, toxicology, environmental effects, aquaculture and seafood quality and it is the first step in the gene banking, to preserve gene materials. The present study we cultured cell of interior pituitary of the Acipenser gueldenstaedtii gland, as a first attempts in IRAN. FISH cell culture has widespread applications in virology, toxicology and as in vitro models in cytogenetic, biomedical, physiological researches. A cell line has been established from the Acipenser gueldenstaedtii interior pituitary and scales have been used to develop primary cell cultures. Recently, successful primary culture of the interior pituitary gland of Acipenser gueldenstaedtii has been developed by explants method. The present study evaluated the potential of several interior pituitary gland from different developmental stages for development of cell cultures using explants method. Pituitary gland from various stages of the adult Acipenser gueldenstaedtii was collected under standard aseptic conditions. Developing gonads from 15-20-year-old male and female sturgeon were collected during late April and early May2007. In all the cases the tissues were pooled in cold PBS antibiotic antimycotic solution (Sigma Chemicals, USA). The tissues were evaluated for attachment, growth and ability to undergo to produce suspension of cells. Primary cultures were initiated from the above tissues according to our earlier procedures, with certain modifications in the sub cultivation procedure. Briefly, tissues were cut into 1 mm3 size fragments which were seeded into 25 cm2 tissue culture flasks. After appropriate semidrying and addition of minimum essential medium (MEM) (Sigma, USA) supplemented with 15% fetal bovine serum (FBS) (Sigma, USA), cell growth was monitored. A seeding density of 1.5 105 cells as determined by a haemocytometer. The results have been showed the Acipenser gueldenstaedtii interior pituitary cells growth in incubator Co2 in 370 C, the cells adapted in this temperature. They were in Lag phase for 10 days, in log phase on 10- 22 days, and in stationary phase on 23- 28 days, after that they died. So we could produce sturgeon growth hormone from fish pituitary cells culture. By this study we can passage the cells on 21th day, for every week. In this way we can produce continued cell culture and store them for gene banking.
Publisher or UniversityIranian Fisheries Science Research Institute
Series : Nr2009.93;