Study on Cry opreservation of Field Sampled Yak Testicular Tissues and Culture of the Isolated Frozen-thawed Spermatogenic Cells
Vitrification freezing was employed to preserve the field sampled testicular tissues of yak.Hematoxy-lin-eosin staining of the frozen testicular tissues indicated that the structure of seminiferous tubules of the testicular tissue was well preserved and a large number of spermatogenic cells in different developmental stages were morpho-logically intact.Trypan blue staining of the frozen-thawed testicular cells showed that the viability rate of the cells was 80.20％.Immunofluorescence staining of the spermatogenic cells and spermatogenic stem cells by the cell marker DDX4 and GF RA1 respectively revealed that the number of spermatogenic stem cells was significantly re-duced after culturing of the frozen-thawed testicular cells for two weeks.RT-qPCR was used to detect the marker gene expressions in the testicular cells in different experimental groups,and the expression of marker genes of sper-matogenic stem cells(Thy1 and UCHL1)was significantly upregulated in the spermatogenic cells cultured for 30 days.Therefore,the structure of seminiferous tubules and spermatogenic cells of yak were well preserved in the vit-rification frozen testicular tissues,which also provided a valuable freezing method to preserve spermatogenic cells of other mammals.
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