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 2

Representative histogram plot for CD11c+ expression on BMDCs. Bone marrow cells were cultured in RPMI 1640 medium with 10% FBS containing low doses of rmGM-CSF (as described in Materials and Methods) and harvested at day 12 for flask culture method and day 5 for EasySep Magnet Positive Selection kit based method. Untreated (sky blue) and low doses of rmGM-CSF treated (pink) BMDCs were stained with PE-labeled anti-CD11c and analyzed by flow cytometry. DCs were gated based on CD11c+ staining. Data were analyzed using Flowjo, LLC, Oregon, USA, software. Images show purity of CD11c+ cells with regard to percentages of adhered CD11c count generated by (a) flask culture method and (c) by EasySep Magnet Positive Selection kit based method and percentages of suspended cell population by (b) flask culture method and (d) by EasySep Magnet Positive Selection kit based method are indicated in the respective histograms .

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