Differentiated cells play an important role in therapeutic strategies for healing patients with tissue injury. Increasing evidence has shown that gold nanoparticles (AuNPs) hold great promise for stimulating cell differentiation. Various stem cells, especially mesenchymal stem cells, can be enhanced by AuNPs. Here, we discuss recent updates on the use of AuNPs for cell differentiation. The impact of AuNPs on the molecular pathways that facilitate cell differentiation is included. We strongly expect that many forms of AuNPs used alone or in combination with scaffolds will be promising for the development of tissue engineering and regenerative medicine.