Cell adhesion is a basic requirement for anchorage-dependent cells to survive on the matrix. It is the first step in a series of cell activities, such as cell diffusion, migration, proliferation, and differentiation. In vivo, cells are surrounded by extracellular matrix (ECM), whose physical and biochemical properties and micromorphology may affect and regulate the function and behavior of cells, causing cell reactions. Cell adhesion is also the basis of communication between cells and the external environment and plays an important role in tissue development. Therefore, the significance of studying cell adhesion in vitro has become increasingly prominent. For instance, in the field of tissue engineering and regenerative medicine, researchers have used artificial surfaces of different materials to simulate the properties of natural ECM, aiming to regulate the behavior of cell adhesion. Understanding the factors that affect cell behavior and how to control cell behavior, including cell adhesion, orientation, migration, and differentiation on artificial surfaces, is essential for materials and life sciences, such as advanced biomedical engineering and tissue engineering. This article reviews various factors affecting cell adhesion as well as the methods and materials often used in investigating cell adhesion.