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A preliminary comparison of dendritic cell maturation by total cellular RNA to total cellular lysate derived from breast cancer stem cells

Phong Minh Le, Tram Thi-Bao Tran, Binh Thanh Vu, Phuc Van Pham.

Abstract
Introduction: Dendritic cells (DCs) have been widely considered as the most potent antigen-presenting cells. As such, DC-based vaccines are regarded as a promising strategy in cancer vaccination and therapy. This study compared the maturation of DCs induced by total cellular RNA and cell lysate (i.e. nucleic acid and protein). Methods: Both total RNA and cell lysate were isolated from breast cancer stem cells (BCSCs). The lysates were used to incubate with monocyte-derived immature DCs. To track the transfection efficiency, the BCSCs were stably transfected with green fluorescent protein (GFP). The maturation of DCs was evaluated by expression of costimulatory molecules including CD40, CD80, and CD86. Transfections were confirmed by evaluating GFP expression in DCs at 24 hours post transfection. Results: The results of this study showed that GFP is expressed in DCs after both total RNA and lysate incubation. The expression of costimulatory molecules (CD40, CD80, and CD86) was significantly higher in RNA-transfected DCs than in cell lysate-primed DCs. Conclusion: Our findings suggest that total RNA primed BCSCs can be a suitable platform for DC-based vaccine therapy of breast cancer.

Key words: Dendritic cells, total cellular RNA, cell lysate, breast cancer stem cell, FuGENE HD, costimulatory molecules



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