“…The most frequently used CSC isolation technique in osteosarcoma was Fluorescent-Activated Cell Sorter (FACS) combined with subsequent culture of the isolated and sorted cells in induction media for maintenance [25,27,31,37,46]. Four articles carried out the sphere formation assay using an induction culture medium [24,28,35,36], one article used FACS only without induction medium [33], two articles used the Magnetic-Activated Cell Sorting (MACS) technique and then induction medium [26,29], one article used MACS without induction medium [34], and finally, the Side Population (SP) technique with subsequent induction medium was used in three articles [13,30,32].…”