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Isolation， Purification and Characterization of Sertoli Cells from Embryonic Chickens
Ke-Jun Cai, Li Liu*, Hai-Lei Ding1, Yi-Xiang Zhang, Zhi-Li Ding, Nian-Ci Zhang
School of Life Sciences, Huzhou Teachers College, Huzhou 313000, China;1College of Animal Science and Technology, Yangzhou University, Yangzhou 225009, China
Abstract: Testes from 18-day-old embryonic chickens were used to isolate seminiferous epithelial cells by sequential two-step enzyme digestion with collagenase and trypsin. During primary culture， highly purified monolayers containing about 90% sertoli cells can be successfully achieved by differential plating and hypotonic treatment. The sertoli cell-enriched cultures were then characterized by staining. The results showed that， no alkaline phosphatase (AKP) staining were found in sertoli cells itself， while another somatic cells， peritubular myoid cells were AKP positive. lipid droplet formation in the cytoplasm and bipolar corpuscula in nucleus were clearly detected after oil red O staining. Acridine orange staining demonstrated that sertoli cells were rich in RNA. Rhodamine 123 staining showed that sertoli cells were rich in mitochondria. Hoechst 33342 staining showed that sertoli cells had a long elliptical shaped nucleus and the long axis of nucleus oriented parallel to the long axis of the cell. Immunofluorescent staining showed that vimentin was expressed in sertoli cell cytoplasm. A simple and reliable method for isolation， purification and characterization of sertoli cells from embryonic chickens has been established.
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