Genomic profiling of human spermatogonial stem cells [WGBS]

To better understand human spermatogonial stem cells (SSCs), we profiled their transciptome and epigenome, which revealed the mechanism how human SSCs regulates their self-renewal versus differentiation dermination, as well as how latent pluripotency is established in human SSCs. Remarkly, we discovered signaling pathways (e.g. LIF, BMP, WNT) that differentially regulated self-renewal vesus differentiation in SSCs. We also discovered that SSCs repress core pluripotent factors (Sox2, Pou5f1 and Nanog) yet activate ancillary factors (e.g. Klf4, Mbd3, Tcf3, Sall4) transcriptionally and epigenetically. Overall design: Using SSEA4+ as self-renewal marker, we isolated self-renewal SSCs by magnetic antibody cell sorting (MACS). Genomic DNA were extracted from isolated cells, and bisulfite converted. Sequencing libraries were then prepared and sequenced on a 125-cycle paired-end run on an Illumina HiSeq 2500 instrument.