BACKGROUND: Human amniotic mesenchymal stem cells are a kind of the ideal seed cells in the field of regenerative medicine. Human amniotic mesenchymal stem cells have shown good application prospect in the treatment of a variety of diseases. Effective control of the key technology of preparation process to ensure that human amniotic mesenchymal stem cells preparations meet the quality requirements is fundament for the safety and effectiveness of clinical application. OBJECTIVE: To describe the key technologies of human amniotic mesenchymal stem cells preparation and quality control, analyze the existing problems and solutions, and summarize the key quality control and technical standards. METHODS: CNKI, Wanfang, and PubMed databases were retrieved by using computer to search relevant articles published from January 2010 to March 2021 with key words of “mesenchymal stem cells, cell culture, biological characteristic, quality control, microbiology contamination” in Chinese and English, separately. Finally, 47 articles were included for analysis. RESULTS AND CONCLUSION: It is the key to quality control and ensure safety in clinic, for example, (1) scientific selection of donors during the preparation of clinical-grade human amniotic mesenchymal stem cells; stable and standardized collection methods; optimization of cell separation, culture, and in vitro expansion techniques, standardization of experimental operations; monitoring of cell preparation laboratory environment, and cell preparation quality detection technology and so on. (2) The establishment and application of a serum-free culture system are conducive to ensuring the quality of preparations. (3) Maintaining the biological characteristics and functional activity of human amniotic mesenchymal stem cells, as well as human amniotic mesenchymal stem cells tumorigenicity, aging, and abnormal immune response meet the release and review inspection quality standards. These can ensure that human amniotic mesenchymal stem cells preparations exert good clinical effects and avoid adverse reactions to the greatest extent. [ABSTRACT FROM AUTHOR]