Chirality is a fundamental feature in biology, from the molecular to the organismal level. An animal has chirality in the left–right asymmetric structure and function of its body. In general, chirality occurring at the molecular and organ/organism scales has been studied separately. However, recently, chirality was found at the cellular level in various species. This “cell chirality” can serve as a link between molecular chirality and that of an organ or animal. Cell chirality is observed in the structure, motility, and cytoplasmic dynamics of cells and the mechanisms of cell chirality formation are beginning to be understood. In all cases studied so far, proteins that interact chirally with F-actin, such as formin and myosin I, play essential roles in cell chirality formation or the switching of a cell’s enantiomorphic state. Thus, the chirality of F-actin may represent the ultimate origin of cell chirality. Links between cell chirality and left–right body asymmetry are also starting to be revealed in various animal species. In this review, the mechanisms of cell chirality formation and its roles in left–right asymmetric development are discussed, with a focus on the fruit fly Drosophila, in which many of the pioneering studies were conducted.