Precartilaginous stem cells (PSCs) could self-renew or differentiate into chondrocytes to promote bone growth. In the current study, we aim to understand the role of transforming growth factor-beta 1 (TGF-beta 1) in precartilaginous stem cell (PSC) proliferation, and to study the underlying mechanisms. We successfully purified and primary-cultured PSCs from the neonate mice' perichondrial mesenchyme, and their phenotype was confirmed by the PSC marker fibroblast growth factor receptor-3 (FGFR-3) overexpression. We found that TGF-beta 1 induced Akt-glycogen synthase kinase-3 beta (GSK3 beta) phosphorylation and beta-catenin nuclear translocation in the mouse PSCs, which was almost blocked by TGF-beta receptor-II (TGFRII) shRNA knockdown. Further, perifosine and MK-2206, two Akt-specific inhibitors, suppressed TGF-beta 1-induced GSK3 beta phosphorylation and beta-catenin nuclear translocation. Akt inhibitors, as well as beta-catenin shRNA knockdown largely inhibited TGF-beta 1-stimulated cyclin D1/c-myc gene transcription and mouse PSC proliferation. Based on these results, we suggest that TGF-beta 1 induces Akt activation to promote beta-catenin nuclear accumulation, which then regulates cyclin D1/c-myc gene transcription to eventually promote mouse PSC proliferation.