|Krasnec, Katina - Us Department Of Agriculture (USDA)|
Submitted to: Poultry Science
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 4/24/2018
Publication Date: 6/4/2018
Citation: Talbot, N.C., Krasnec, K.V., Garrett, W.M., Shannon, A.E., Long, J.A. 2018. Finite cell lines of turkey sperm storage tubule cells: ultrastructure and protein analysis. Poultry Science. https://doi.org/10.3382/ps/pey208.
DOI: https://doi.org/10.3382/ps/pey208 Interpretive Summary: The turkey, like chickens and other birds, has the ability to store sperm for several weeks after mating with the male turkey. The turkey hen has specialized little sacs, called sperm storage tubules or SSTs, at the top of its vagina that somehow store the sperm for all that time. How this is accomplished by the SSTs is being studied in the hope that better methods might be developed for storing turkey sperm for use in the turkey production business. Currently, without the ability to store sperm for more than a few hours, turkey production is labor intensive, requiring the collection of fresh sperm from male turkeys every day. In the current report, research was conducted to grow turkey SST cells in vitro, i.e., in petri dishes, so that experiments on how SSTs are able to store sperm can be performed. In vitro cell cultures of the turkey sperm SST cells were established from freshly obtained uterovaginal junction (UVJ) tissue. Three independent cell lines, SST-1, -2 and -3, were for a period of 2-3 months. In vitro growth conditions were investigated, and the characteristics of the SST cell cultures were compared to the SST cells in the hen's body to see how alike they were. It was discovered that the SST cells grown in the dish were different from the SST cell in the body because those in the dish produced large amounts of mucus. More research on culture turkey SST cells will be necessary to achieve a better in vitro model of the turkey hen's SST cells as they exist in the body.
Technical Abstract: Cell lines of turkey sperm storage tubule (SST) epithelial cells were established. Turkey SSTs were dissected from freshly obtained uterovaginal junction (UVJ) tissue and placed in explants culture on various substrates and media. Primary cultures of SST epithelium only survived and grew from SST explants that were cultured on inactivate Sandoz inbred strain, thioguanine- and ouabain-resistance (STO) mouse feeder-cell layers in 12% fetal bovine serum-supplemented Dulbecco's Modified Eagle Medium mixed 1:1 with F12 nutrient mixture. Three independent primary colonies gave rise to three finite cell lines, SST-1, -2 and -3, which were continuously cultured for 8-16 passages at 1:3 passage ratios over a period of 3-4 months. The cells were passaged by pretreatment with Y27632 and dissociation with Accutase. The SST cells grew as tightly knit monolayers on top of the feeder-cells at a slow rate (~96 h doubling time) at a medium pH of ~6.9. Lipid vacuoles were visible by light microscopy in the cells particularly at the periphery of growth. Transmission electron microscopy revealed the cells to be a polarized epithelium with apical microvilli and to have lateral tight-junction-like unions and associated desmosomes. Numerous secretory vesicles filled the upper portion of the cells' cytoplasm, and nuclei and other major organelles such as mitochondria, rough endoplasmic reticulum, and Golgi apparatus were distributed somewhat lower in the cytoplasm. The secretory vesicles resembled mucin secretory vesicles. Proteomic analysis by mass spectroscopy of the conditioned medium of the cells, and of the cells themselves, showed the cell lines did not secrete large amounts of any particular protein, and the analysis confirmed their epithelial character. In conclusion, the SST-derived cell lines resembled the mucus-secreting cells found in the epithelium lining UVJ of the turkey's reproductive tract.