Fractionation of human spermatogenic cells using STA-PUT gravity sedimentation and their miRNA profiling

• DOI: 10.1038/srep08084
• Publication Date: 2015-01-30T13:56:59Z
• Authors: Yun Liu; Minghui Niu; Chencheng Yao; Yanan Hai; Qingqing Yuan; Yang Liu; Ying Guo; Zheng Li; Zuping He
• Keywords: Spermiogenesis

Abstract Human spermatogenic cells have not yet been isolated and notably, their global miRNA profiles remain unknown. Here we effectively human spermatogonia, pachytene spermatocytes round spermatids using STA-PUT velocity sedimentation. RT-PCR, immunocytochemistry meiosis spread assays revealed that the purities of were 90% viability these was over 98%. MiRNA microarrays showed distinct among spermatids. Thirty-two miRNAs significantly up-regulated whereas 78 down-regulated between spermatogonia spermatocytes, suggesting are involved in mitosis, respectively. In total, 144 while 29 spermatids, reflecting potential roles mediating spermiogenesis. A number novel binding targets further identified various softwares verified by real-time PCR. Our ability isolating unveiling signatures could provide small RNA regulatory mechanisms three phases spermatogenesis offer new for treatment male infertility.