Cell adhesion and its role during cell spreading and motility are central to normal development and homeostasis, including its effects on immune response and wound repair and tissue regeneration. Disruption of cell adhesion impacts not only the healing process but promotes tumor invasion and metastasis. A family of intracellular, limited proteases, the calpains, has recently been shown to be a key molecular control point in attachment of cells to the surrounding matrix. Herein, the two main and ubiquitously expressed calpain isoforms will be introduced as to their modes of regulation and the current status of research will be discussed as to how these calpains might function in the biophysical process of adhesion and biological cellular responses of spreading and motility.