ABSTRACT
Objective To isolate carboplatin-resistant human ovarian cancer stem-like cells and provide a cellular model for the development and screening of second-line drugs for carboplatin-resistant ovarian cancer.Methods Primary ovarian cancer cells were isolated and cultured from ovarian cancer tissues and ascites,and ovarian cancer stem-like cell spheres were isolated from serum-free suspension culture directly or after carboplatin induction.The expression of stem cell markers was evaluated by immunofluorescent assay or flow cytometry.Results The ovarian cancer stem-like cell spheres were isolated successfully from both ovarian cancer tissues and ascites and expressed high levels of stem cell markers CD133,ALDH1,ABCG2,and CD24.The abundance of CD24+ cells in the ovarian cancer stem cells was significantly increased upon short-term induction with carboplatin.Condusion Ovarian cancer stem-like cells can be enriched and purified by short-term induction with carboplatin.This cell model can be used in research on chemoresistant ovarian cancer in the future.
ABSTRACT
ABSTRACT Lentiviral vector-mediated gene transfer offers several advantages over other gene delivery vectors when considering gene and cell therapy applications. However, using these therapies in clinical applications involves large-scale vector production in an efficient and cost-effective manner. Here we describe a high yield production of a lentivirus encoding recombinant factor VIII in a scalable and GMP-compliant culture system, based on serum free suspension cultures and transient transfection with an inexpensive reagent, polyethylenimine (PEI), reaching a total viral yield of 2.48x108 particles.