In order to prepare acrylamide copolymers with non-toxicity for environment-friendly materials, a series of zwitterionic acrylamide copolymers with varying acrylamide content were synthesized with DMAPS (3-dimethyl (methacryloyloxyethyl) ammonium propane sulfonate) and acrylamide via free radical polymerization. The structural properties and cytotoxicity are carefully investigated. The results demonstrate that DMAPS-co-acrylamide copolymers contain DMAPS and acrylamide segments that form a matrix consisting of strong electrostatic interactions as well as strong hydrogen bonding. DMAPS segments exhibit negligible influence on the glass phase transition behavior, yet affect the composite's water-contact angle. The optimum hydrophilic properties are obtained by adjusting the acrylamide content to 50 wt%. All DMAPS-co-acrylamide copolymers are measured to be non-toxic. The DMAPS segments allow for suitable contribution to the biocompatibility of the synthesized copolymer, in which copolymers containing higher DMAPS content demonstrate better biocompatibility.