The majority of staphylococci produce biofilm on medical devices, which is the main mechanism to infect humans. Staphylococcal biofilms attach to abiotic or biotic surfaces, forming aggregates and protecting themselves against the immune system and the antimicrobial compounds of the host. Few studies on biofilm formation mechanism in Staphylococcus epidermidis and other coagulase-negative staphylococci (CNS) have been performed; however, there is a great interest in studying and controlling biofilm formation of this genus. This chapter exhibits the state of the art on biofilm formation in S. epidermidis and other staphylococcal species. The main goal of this chapter is to recognize the importance of biofilm formation in Staphylococcus. The participating molecules in staphylococcal biofilm formation are described. Currently, biofilm producer strains of Staphylococcus and mainly CNS have been frequently isolated at hospitals, causing significant economic losses. This chapter includes promising solutions in order to prevent medical device-associated infections, as the development of medical devices possessing anti-biofilm materials or surfaces that act against the adhesion or viability of the microorganisms.