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目的探讨人原代卵巢癌细胞的提取及其干细胞筛选的具体方法。方法用含EDTA的胰蛋白酶消化新鲜无菌的卵巢癌组织,过滤后用Percoll细胞分离液分离出卵巢癌细胞,并用特定配制的干细胞选择性培养基培养卵巢癌细胞,培养21d筛选出悬浮生长的卵巢癌干细胞,并运用Q-PCR,流式细胞术验证干细胞标志。结果运用干细胞选择性培养基培养卵巢癌细胞,能够筛选出表面分子CD44和CD117阳性的绝大多数卵巢癌干细胞,且其干性基因OCT-4、Twist、Snail、Naong和Slug的表达水平显著升高。结论运用特定配制的干细胞选择性培养基能够筛选出绝大多数的卵巢癌干细胞,这是一种较为简便的获得干细胞的方法,对研究干细胞的生物学特征、筛选化疗药物及指导临床治疗具有重大意义。
Objective To explore the method of extracting human primary ovarian cancer cells and screening their stem cells. Methods Fresh sterile ovarian cancer tissues were digested with trypsin containing EDTA, and ovarian cancer cells were isolated by Percoll cell separation after filtration. The ovarian cancer cells were cultured with specially selected stem cell selective medium and cultured for 21 days. Ovarian cancer stem cells, and the use of Q-PCR, flow cytometry to verify stem cell markers. Results The majority of ovarian cancer stem cells positive for surface molecules CD44 and CD117 were screened by culturing ovarian cancer cells with stem cell selective medium and the expression levels of OCT-4, Twist, Snail, Naong and Slug were significantly increased high. Conclusion It is a simple and convenient method to obtain stem cells that can screen most of the ovarian cancer stem cells using specially formulated stem cell selective medium. It is significant to study the biological characteristics of stem cells, screen chemotherapeutic drugs and guide the clinical treatment significance.