Abstract: The aim of this study was to investigate the capability of spermatogonial stem cells(SSCs) differentiating into osteoblasts, neuron-like cells,and adipocytes in vitro. In present study, SSCs were isolated from testis of chicken embryo on 18^th-20^th hatching days, then cultured and subcultured to the 3rd passage in vitro. ① SSCs were Orientate-induced into osteoblasts by medium consisting of desamethason, glycerol 2-phosphate disodium salt hydrate and vitamin C, and differentiated cell were examined with Von Kossa’s stain,cytochemistry staining and immunohistochemistry stain. ②SSCs were Orientate-induced into neuron-like cells in the medium containing RA and IBMX. The differentiated cells were identified by toluidine blue and immunohistochemistry stain. ③SSCs were induced into adipocytes in the medium consisting of desamethason, insulin and IBMX. The differentiated cells were then identified by oil red-O and specific express PPARγ. The results showed that 80% SSCs were induced into osteoblasts during 15-21 d after cultured in induction medium. The osteoblasts formed Von Kossa stain positive nodules. Dark granules were found in the cell, suggesting the deposition of mineralized materials. The osteoblasts showed brown or deep dark after alkaline phosphatase stain and showed CollagenⅠpositive. About 85% cells was toluidine blue positive after 7 d when SSCs were induced into neuron-like cells. About 85% oil red-O positive cell formed during 14-21 d when SSCs were induced into adipocytes. The induced cells were highly expressed PPARγ. Therefore, we concluded that chicken SSCs have the capability of multi-differentiation in vitro.