Isolation and Purification of Murine Male Germ Cells

Seminiferous tubules of the male testis contain somatic cells (Sertoli and Leydig cells) and germ cells at different stages of spermatogenesis (spermatogonia, spermatocytes and spermatids). Germ cells at different stages of differentiation migrate toward the central lumen via cell junctions formed by Sertoli cells. The protocol described herein consists of the dissection and decapsulation of the testes, disruption of the structure of the seminiferous tubules, and the breaking of cell junctions to release all the cells in suspension. Germ cells are then separated from Sertoli cells by overnight plating of the suspension on plastic to which the germ cells preferentially adhere. And finally, a BSA gradient allows a high-purity separation of the various types of germ cells according to size.