Culture and Expansion of Primary Undifferentiated Spermatogonial Stem Cells

This protocol describes a culture method for supporting the maintenance and expansion of a primary mouse undifferentiated spermatogonial population containing spermatogonial stem cells (SSCs). The doubling time for SSCs in culture is relatively slow. Once established, SSC lines are split 1:2 to 1:4 every 7 d. Therefore, the time required to generate sufficient SSCs for experimentation can be considerable and requires careful planning.