Abstract: Cancer stem cell hypothesis has become a new theory. According to this theory, cancer stem cells play a decisive role in carcinogenesis, progression, and metastasis. This theory offered new ideas and strategies for cancer treatment. However, only a small fraction of cells in a tumor were cancer stem cells. Therefore, the identification and enrichment of these cells are significant challenges for numerous researchers conducting further study. Micro-sphere culture has been an effective tumor stem cell enrichment technology in recent years. The application of this technique makes the cancer stem cell research process easy. In addition, it makes the evaluation of tumor progression and prognosis of patients in clinical possible. This article will focus on the applications of this technology, and microsystem integration will be described in detail.