Establishing Induced Pluripotent Stem Cells With Non-obstructed Azoospermia Syndrome

Infertility accounts for15%of human population, in which40%is due to male reproductive factors [1]. In cases of male infertility, non-obstructive azoospermia (non-obstructed azoospermia, NOA) accounts from10%[2,3]. It is currently very challenging to investigate the pathology and molecular mechanisms of non-obstructive azoospermia (NOA) in vitro. So, induced pluripotent stem cell (iPS) obtained from terminally differentiated cells of a NOA patient,may overcome this technical problems. In this study, we isolated primary fibroblasts and epithelial cells from5NOA-patients and established more than70iPS cell lines. These iPS cell lines induced them into pluripotent stem cells with four factors OCT4, SOX2, KLF4and c-Myc. We have obtained a total of60iPS cell lines from5NOA pathological samples of NOA-patients. We have character these iPS cell lines. They share the same characteristics with human embryonic stem cells in morphology,AP staining,and marker expression. In addition, we confirmed that these iPS cell lines come from patients via the micro-satellite (short term repeat, STR) technology and other molecular biology tools, Specifically, we found that2samples, P2618,andP2514,have micro deletion in the infertility factor (azoospermia factors, AZF) c area in genome, which directly led to the reduced copies and expression level of a key germ cell specific gene, DAZ[4], in DAZ family. In addition, accumulating evidences suggest that gene mutations or nucleotide polymorphism result in serious defects of spermatogenesis. The single nucleotide polymorphism (single nucleotide polymorphisms, SNP) screening was thus performed on these samples, and interestingly,we found that sample P2656hade polymorphisms at two SNP sites,rs7192and rs2477686, which have high correlation with NOA syndrome [5,6]. Currently, only a few studies reported a correlation about SNP sites and NOA syndrome and have not been experimentally verified Therefore, our NOA-iPS lines will provide a good platform for this study. Taken together, our research provide an useful model for further exploring the pathogenesis of non-obstructive azoospermia in vitro.