Objective To review the latest development of amniotic fluid-derived stem cells (AFSCs) in regenerative medicine, and to discuss issues related to the studies in the field of AFSCs. Methods The recent articles about AFSCs were extensively reviewed. The important knowledge of AFSCs was introduced in the field of regenerative medicine, and the basic and clinical researches of AFSCs were summarized and discussed. Results Currently, it is confirmed that AFSCs have a multi-directional differentiation capacity, therefore, they have a wide application prospect in regenerative medicine, anti-tumor, and other fields. Conclusion AFSCs will become one of the ideal seed cells in the field of regenerative medicine with extensive research value because of the advantages of easy amniotic fluid sampling, little maternal and child trauma, no tumorigenesis, and no ethical restrictions.