Isolation,Culture and Identification of Sertoli Cells from Dairy Goat
In this study,the Sertoli cells(SCs)were isolated from 2-month-old Shannon dairy goats,and the isolation efficiency of three enzyme digestion methods(trypsin digestion,two-step enzyme digestion,and two-step mixed enzyme digestion)were compared.The morphological and molecular biological techniques were used to characterize and evaluate the culture effects.The results showed that the number,viability and proliferation of cells obtained by two-step enzyme digestion and two-step mixed enzyme digestion were signifi-cantly higher than those obtained by pancreatic enzyme digestion.Morphological observation showed that the cells isolated by two-step enzyme digestion method had typical SCs morphology and could be stably passaged to the fifth generation.HE staining showed bipolar corpuscula in the nucleus,and oil red O staining showed lipid droplets with different sizes around the nucleus.Immunofluorescence staining showed that the positive rate of SCs specific protein WT1 and Vimentin antibodies was more than 96％.The ultrastructural observation showed that the cell structure was intact,the organelles were abundant,and the nucleus was irregular in shape,which was consistent with the ultrastructural characteristics of SCs.In conclusion,the two-step enzyme digestion method was simple,low-cost,and could isolate a large number of SCs with good viability and purity,which could be stably cultured in vitro,so it was the best method for isolating SCs from dairy goats,and could pro-vide a good cytological model for the in-depth study of male reproductive function of dairy goats.
Dairy goatSertoli cellsIsolation and culture methodsIdentificationUltrastructure
万方数据
维普
