The testicular cells of Landrace piglets aged from 1 to 5 days were isolated by two-step enzymatic digestion method. The Percoll discontinuous density gradients method was used to purify the porcine spermatogonial stem cells (PSSCs). The effects of glial cell-line derived neurotrophic factor (GDNF) and leukemia inhibitory factor (LIF) on PSSCs proliferation were investigated by MTT method. The results showed that the method for PSSCs isolation and purification were effective. By means of MTT assay, the results indicated that LIF and GDNF factor could promote PSSCs proliferation, and the combination of 20 ng·mL^-1 GDNF and 1 000 U·mL^-1 LIF showed the strongest effects.