Two-dimensional materials (2DMs) with excellent mechanical, thermal, optical, and catalytic properties have attracted a great deal of attention in recent years. Chemical vapor deposition (CVD) is an important method to realize the synthesis of high-quality 2DMs. In the growth of 2DMs through the CVD method, the substrates play an important role and can greatly affect the lateral size, composition, thickness, orientation, and crystal quality of 2DMs. In this review, we first introduce the growth mechanism and the key parameters in the CVD system for the synthesis of 2DMs. Then, the unique physical and chemical properties, advantages, and disadvantages of various substrates used in the CVD technique are summarized. Finally, the opportunities and challenges about the use of the substrate in the CVD process in the future are discussed.