Spermatogonial stem cells (SSCs) developed into sperms through spermatogenesis have been utilized as a useful tool in the field of regenerative medicine and infertility. However, a small number of highly qualified SSCs are resided in the seminiferous tubule of testis, resulted in developing effective in-vitro culture system of SSCs for solving simultaneously quantitative and qualitative problems. Presently, SSCs can be enriched on testicular stromal cells (TSCs), but there are no systematic researches about TSC culture. Therefore, we tried to optimize culture condition of TSCs derived from mouse with different strains. For these, proliferation and viability were measured and compared by culturing ICR outbred or DBA/2 inbred mouse-derived TSCs at 35 or $37^{circ}C$. In case of ICR strain, primary TSCs cultured at $37^{circ}C$ showed significantly higher proliferation and viability than those at $35^{circ}C$ and significant increase of proliferation and viability in sub-passaged TSCs was detected in the $35^{circ}C$ culture condition. Moreover, sub-passage of primary TSCs at $35^{circ}C$ induced no significant effects on proliferation and viability. In contrast, in case of DBA/2 strain, significantly improved proliferation were detected in the primary TSCs cultured at $35^{circ}C$, which showed no significant difference in the viability, compared to those at $37^{circ}C$. Furthermore, sub-passaged TSCs cultured at $37^{circ}C$ showed no significant differences in proliferation and viability, compared to those at $35^{circ}C$. However, with significant decrease of proliferation induced by sub-passage of primary TSCs at $35^{circ}C$, no significant effects on proliferation and viability were resulted from sub-passage of primary TSCs at $37^{circ}C$. From these results, culture temperature of primary TSCs derived from outbred and inbred strain of mouse could be separately optimized in primary culture and subculture.