This article describes a protocol for isolating human spermatogonial stem cells (hSSCs) from human testis tissues, which can be utilized for further characterization. Human testis tissues are first subjected to mechanical separation, enzymatic digestion, and physical filtering, yielding a single testicular cell suspension. SSCs or other cell types of interest can be sorted out via different enrichment approaches using selected cell surface markers (e.g., magnetic-activated cell sorting, MACS). Those enriched cells can further be used for different purposes, including cell culture, RNA sequencing, and other genomics-based profiling, which allow for further characterization of their properties. © 2019 by John Wiley & Sons, Inc.
Keywords: cell sorting; hSSCs; human spermatogonial stem cells; single-cell isolation.
© 2019 John Wiley & Sons, Inc.