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Fig. 6 | Stem Cell Research & Therapy

Fig. 6

From: Bone marrow mesenchymal stem cells tune the differentiation of myeloid-derived suppressor cells in bleomycin-induced lung injury

Fig. 6

BMSC tune the differentiation of MDSC in vitro. Gr-1+CD11b+ cells (5 × 104 cells/well) isolated from spleens of naïve C57BL/6 mice (n = 3) were cultured alone or in the presence of NIH-3 T3 or BMSC (1 × 104 cells/well). A total of 24, 48, and 72 h after coculture, floating cells were collected and numerated. a Seventy-two hours after coculture, the phenotype of floating cells was analyzed by flow cytometry (FCM). Representative FCM data are shown. b The absolute number of Gr-1+CD11b+ cells and (c) the variations of Gr-1HighCD11b+/Gr-1LowCD11b+ were calculated. d-f BMSC and Gr-1+CD11b+ cells were co-cultured and transwell inserts were used in some wells to prevent cell-cell contact. d Seventy-two hours after coculture, the phenotype of floating cells was analyzed by flow cytometry (FCM). Representative FCM data are shown. e The absolute number of Gr-1+CD11b+ cells and (f) the variations of Gr-1HighCD11b+/Gr-1LowCD11b+ were calculated. g The concentration of M-CSF was measured in the supernatant using ELISA. h Gr-1+CD11b+ cells isolated from spleen of naïve C57BL/6 mice were cultured with exogenous recombinant mouse M-CSF. The variations of Gr-1HighCD11b+/Gr-1LowCD11b+ were calculated. i-j Gr-1+CD11b+ cells isolated from spleens of naïve C57BL/6 mice were cocultured with BMSC transfected with either control siRNA or siM-CSF (n = 3). i The absolute number of Gr-1+CD11b+ cells and (j) the variations of Gr-1HighCD11b+/Gr-1LowCD11b+ were calculated. Data presented are representative of two replicated experiments. *P < 0.05, **P < 0.01 and ***P < 0.001. BMSC bone marrow mesenchymal stem cells, mAb monoclonal antibody, W/O without

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