Here, we will for the first time isolate CD133+/CD24+ cells from RCC ACHN cell line using MACS and validate the expression of its stemness-associated makers (CTR2, BCL-2, MDR1, OCT-4, KLF4) and its stem-like characteristics including self-renewal capability, chemoresistance, metastatic potential and tumorigenicity in vivo. The gene discussed is POU5F1; the disease is renal cell carcinoma.