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Abstract To explore the isolation and culture method of duck testis sertoli cells, a total of 40 duck embryos of 24 days old were selected in this experiment. Testis was taken out, and the testis cells were separated by two-step digestion with collagenase IV and trypsin, and then purified by differential adherence and hypotonic treatment. The cells were cultured at 37 ℃, 5% carbon dioxide, and finally high purity sertoli cells were obtained. After staining and identification, rhodamine 123 staining showed that the cells were rich in mitochondria. AKP test showed that 80% of the cells were AKP negative. Acridine orange staining showed that the cytoplasm of cells was rich in RNA. Oil red O staining showed that the cytoplasm of cells contained a large number of fat droplets and bipolar bodies were visible in the nucleus.The results of this experiment showed that high purity sertoli cells could be obtained by combining collagenase IV with trypsin for digestion and purification through differential adherence and hypotonic treatment. Combining rhodamine 123 staining, AKP staining, acridine orange staining and oil red O staining could effectively identify sertoli cells.
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Received: 18 September 2020
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2019, Vol. 32 
