Zinc-based biodegradable metals (BMs) have been developed for biomedical implant materials. However, the cytotoxicity of Zn and its alloys has caused controversy. This work aims to investigate whether Zn and its alloys possess cytotoxic effects and the corresponding influence factors. According to the guidelines of the PRISMA statement, an electronic combined hand search was conducted to retrieve articles published in PubMed, Web of Science, and Scopus (2013.1–2023.2) following the PICOS strategy. Eighty-six eligible articles were included. The quality of the included toxicity studies was assessed utilizing the ToxRTool. Among the included articles, extract tests were performed in 83 studies, and direct contact tests were conducted in 18 studies. According to the results of this review, the cytotoxicity of Zn-based BMs is mainly determined by three factors, namely, Zn-based materials, tested cells, and test system. Notably, Zn and its alloys did not exhibit cytotoxic effects under certain test conditions, but significant heterogeneity existed in the implementation of the cytotoxicity evaluation. Furthermore, there is currently a relatively lower quality of current cytotoxicity evaluation in Zn-based BMs owing to the adoption of nonuniform standards. Establishing a standardized in vitro toxicity assessment system for Zn-based BMs is required for future investigations.