Graphene, as a class of 2D carbon nanomaterial, has attracted tremendous interest in different areas in recent years including biomedicine. The toxicity and behavior of graphene in biological systems are thus important fundamental issues that require significant attention. In this article, the toxicity of graphene is reviewed by describing the behavior of graphene and its derivatives in microorganisms, cells, and animals. Despite certain inconsistencies in several detailed experimental results and hypotheses of toxicity mechanisms, results from numerous reports all agree that the physicochemical properties such as surface functional groups, charges, coatings, sizes, and structural defects of graphene may affect its in vitro/in vivo behavior as well as its toxicity in biological systems. It is hoped that this review article will provide an overview understanding of the impacts, behavior, and toxicology of graphene and its derivatives in various biological systems.