Oogonial stem cell isolation and culture from brow trout (Salmo trutta, Linneaus 1766)

A practical technique for oogonial stem cell isolation and culture of the female Brown trout (Salmo trutta macrostigma, S.t.macrostigma) were elucided in this thesis. Initially, appropriate age, size and weight of the female S.t. macrostigma for oogonial stem cell isolation and culture were identified as 2,5-3 month old, 14,65±1,66 cm, 28,22±7.79 g respectively. Secondly, morphological and histological ovary conditions were assessed. Finally, isolation and culture conditions for oogonial stem cells of S.t. macrostigma were optimized. The highest oogonial stem cells were measured in the perinucleolar stage of the ovary. Survival rate of oogonia and mitotic activities in L-15 culture media, with 5% serum of S.t. macrostigma (Culture media pH, 7,65; temperature 20ºC) were developed. Density of oogonial stem cells were measured as 5,4*105 ±2,6*105 cells/ml. Whereas, oogonial volume was recorded as 13,5 ml. In the thesis, germ line chimera was developed for S.t. macrostigma in order to be used in surrogate reproduction technologies and gene transfer systems. After culture of oogonial stem cells it was suggested that 2,7*106 ± 1,3*106 cells/ ml cell count should be used in transplantation technologies.