Optimization of Ex Vivo Murine Bone Marrow Derived Immature Dendritic Cells: A Comparative Analysis of Flask Culture Method and Mouse CD11c Positive Selection Kit Method

Figure 4

Expression of surface markers of adhered and suspended rmGM- BMDCs generated in the absence of LPS. FACS surface profile of surface markers BM-DCs from BALB/c mouse at day 12 for flask culture method and day 5 for EasySep Magnet Positive Selection kit based method. Bone marrow cells were cultured in RPMI 1640 medium with 10% FBS containing low doses of rmGM-CSF (as describe in Materials and Methods) and harvested, respectively, for above-mentioned methods. Untreated blank (sky blue) and low doses of rmGM-CSF treated (pink) adhered and suspended BMDCs were stained and analyzed by flow cytometry. Images show representative histograms of immature BM-DCs expressing cell population of adhered MHCII+ (a), suspended MHCII+ (c), adhered CD86+ (e), suspended CD86+ (g), adhered CD40+ (i), and suspended CD40+ (k) costimulatory molecules generated by flask culture method and adhered MHCII+ (b), suspended MHCII+ (d), adhered CD86+ (f), suspended CD86+ (h), adhered CD40+ (j), and suspended CD40+ (l) costimulatory molecules by EasySep Magnet Positive Selection kit based method.

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