Cell growing behavior is significantly dependent on the surface chemistry of materials. SU-8 as an epoxy-based negative photoresist is commonly used for fabricating patterned layers in lab-on-a-chip devices. As a hydrophobic material, SU-8 substrate is not favorable for cell culture, and cell attachment on native SU-8 is limited attributed to poor surface biocompatibility. Although physical adsorption of proteins could enhance the cell adhesion, the effect is not durable. In this work, SU-8 surface chemistry is modified by immobilizing fibronectin (FN) and collagen type I (COL I) covalently using (3-aminopropyl)triethoxysilane (APTES) and cross-linker glutaraldehyde (GA) to increase surface biofunctionality. The effectiveness of this surface treatment to improve the adhesion and viability of mesenchymal stem cells (MSCs) is investigated. It is found that the wettability of SU-8 surface can be significantly increased by this chemical modification. In addition, the spreading area of MSCs increases on the SU-8 surfaces with covalently conjugated matrix proteins, as compared to other unmodified SU-8 surface or those coated with proteins simply by physical adsorption. Furthermore, cell proliferation is dramatically enhanced on the SU-8 surfaces modified under the proposed scheme. Therefore, SU-8 surface modification with covalently bound matrix proteins assisted by APTES+GA provides a highly biocompatible interface for the enhanced adhesion, spreading, and proliferation of MSCs.