In this paper, the in vitro biocompatibility of graphene film (GF) with osteoblasts was evalu-ated through cell adhesion, viability, alkaline phosphatase activity, F-actin and vinculin ex-pressions, versus graphite paper as a reference material. The results showed that MG-63 cells exhibited stronger cell adhesion, better proliferation and viability on GF, and osteoblasts cultured on GF exhibited vinculin __EXPRESSION__ throughout the cell body. The rougher and wrinkled surface morphology, higher elastic modulus and easy out-of-plane deformation associated with GF were considered to promote cell adhesion. Also, the biomineralization of GF was assessed by soaking in simulated body fluid, and the GF exhibited enhanced miner-alization ability in terms of mineral deposition, which almost pervaded the entire GF surface. Our results suggest that graphene promotes cell adhesion, activity and the formation of bone-like apatite. This research is expected to facilitate a better understanding of graphene-cell interactions and potential applications of graphene as a promising toughening nanofiller in bioceramics used in load-bearing implants.