Single-cell Gene Expression Analysis Reveals Transcriptional Diversity among Human Spermatogonia. Homo sapiens

Two individual human OCT4-positive spermatogonia were selected for global transcriptional capture using shallow RNA-seq. Successful cDNA synthesis and amplification was confirmed as reads mapped primarily to exons. It has to be noted though, that a comparision of cell to cell data is not possible as coverage of the whole transcriptome was too low, approximately <1.0x coverage. Nonetheless, RNA-seq data provided first information of highly abundant transcripts in individual human spermatogonia. Spermatogonia-specific expression of selected candidate genes with high RPKM values including DDX5, EEF1A1 and TSPY1 was confirmed by immunohistochemistry. Overall design: Human testicular biopsies with qualitatively normal spermatogenesis were used to establish human germ cell cultures. Spermatogonia were selected based on morphological characteristics via micromanipulation.