Abstract: In recent years, the probability of discovering new chemical structures and bioactive substances continues to decline. In order to overcome the disadvantages of traditional culture methods, new methods for microbial culture were developed, including co-culture, highthroughput culture, in situ culture, and soil substrate membrane system (SSMS) methods. This review mainly summarizes the selected cases of the aforementioned four methods in recent years, hoping to provide references for future microbial culture studies, with the purposes to discover more new natural products.