Isolation and culture of cancer stem cells from ovarian cancer tissues and malignant ascites / 肿瘤

ABSTRACT

Objective: To isolate and culture tumor stem cells from ovarian cancer tissues and malignant ascites, and to analyze the cellular phenotype and drug sensitivity. Methods: The primary tumor tissues and the malignant ascites samples from one patient with ovarian adenocarcinoma were collected, and the formations of float-growing spheres was developed with serum-free medium culture. The expressions of stem cell markers were evaluated by flow cytometry (FCM) and immunofluorescent staining. The sensibilities of sphere cells and differentiated cells to paclitaxol and carboplatin were measured by in vitro drug sensitivity assay. Results: The ovarian cancer cells in both tumor tissues and malignant ascites displayed an ability to generate spheres cultured in serum-free medium. These sphere-forming cells exhibited the phenotype of CD44+CD24- and also expressed stem cell markers including Oct4, Nanog and Sox2. These sphere cells from tumor tissues and malignant ascites became differentiated after culture in serum-containing medium for 3 weeks, and the fractions of CD44+ and CD24- cells reduced by 47% and 3%, respectively. The differentiated cells showed increased sensitivities to paclitaxol and carboplatin as compared with the sphere cells (P<0.01). Conclusion: The ovarian cancer stem cells exhibit the phenotype of CD44+CD24-, which is characterized by expressions of several stem cell markers including Oct4 and a high level of drug resistance. Copyright© 2011 by TUMOR.