Neonatal Testicular Gonocytes Isolation and Processing for Immunocytochemical Analysis

In recent years, an increasing interest has emerged at understanding how spermatogonial stem cells (SSCs) arise from their precursor cells, the gonocytes. The identification of factors acting directly on gonocytes rather than on the surrounding somatic cells and the study of genes and signaling pathways intrinsic to gonocytes require their isolation from Sertoli and peritubular cells. The present article describes a simple method developed to isolate rat neonatal gonocytes, allowing for the study of their proliferation and differentiation to SSCs. We also present immunocytochemical methods that can be used to study protein expression changes and proliferation in gonocytes.