Journal: Cell stem cell
Article Title: Gli1+ mesenchymal stromal cells are a key driver of bone marrow fibrosis and an important cellular therapeutic target
Figure Lengend Snippet: Gli1 + cells reside in the perivascular and endosteal bone marrow niche in human and mice (A ) Representative images of bone marrows (BM) from bigenic Gli1CreER t2 ;tdTomato mice 10 days after tamoxifen application (3x10mg p.o.). Brightfield (BF) and Fetuin A stained images. Scale bars 50μm (B-C) Representative images and Z-Stack analyses of the perivascular BM niche. Scale bars 50μm. (D) Distribution of Gli1 + cells in the endosteal (endost), perisinusoidal (perisin) and periarteriolar (periart) niche; mean±SEM. (E-H) CD105, NG2 and Nestin expression by Gli1 + cells in the endosteal, perisinusoidal and periarteriolar niche; mean±SEM; arrows indicate co-expression in Gli1 + cells; arrowheads indicate no-expression in Gli1 + cells; scale bars 50μm in E-F, 25μm in G-H; mean±SEM. (I) Representative Z-Stack images and quantification of Gli1 + cell innervation by tyrosine hydroxylase (TyrHydr)-positive sympathic nerve fibres. Scale bars 25μm; mean±SEM. (J) Representative Z-Stack image and quantification of Gli1 + cells adjacent to glial fibrillary acidic protein (GFAP)-expressing glia cells. Scale bars 25μm. (K) Surface expression pattern of tdTomato + cells from bigenic Gli1CreER t2 ; tdTomato mice as assessed by flow cytometry; mean±SEM. ( L ) Representative images of human bone marrows stained for Gli1. Arrows indicate Gli1 expressing cells. Scale bars: 50μm . (M) Surface expression profile of human Gli1 + .
Article Snippet: Cells were labeled with the following monoclonal, directly fluorochrome conjugated antibodies: anti-mouse: Ter119 (eBioscience efluor 450), Gr1 eBioscience (efluor 450), CD11b (eBioscience, efluor 450), B220 (efluor 450 eBioscience), CD3 (efluor 450 eBioscience), CD150 (PeCy7 BioLegend), CD48 (APCCy7 Biolegend), CD34 (Alexa Fluor 700 eBioscience), ckit (APC eBioscience), Sca1 (PE, eBioscience), Gr1 (Alexa Fluor 700, eBioscience), CD11b (APC, eBioscience), CD3 (PE, eBioscience), CD19 (PeCy7 eBioscience), Ter119 (FITC, eBioscience), CD31 (APC, Biolegend), CD34 (FITC, eBioscience), CD29 (APC, eBioscience), Sca1 (APC-Cy7, Biolegend), CD44 (PE-Cy7 eBioscience); CD105 (PE-Cy7, Biolegend), CD73 (APC, Biolegend), PDGFRα (APC, Biolegend), CD31 (APC, Biolegend), CD34 (FITC, eBioscience), CD29 (APC, eBioscience), Sca1 (APC-Cy7, Biolegend), CD44 (PE-Cy7 eBioscience); CD105 (PE-Cy7, Biolegend), CD73 (APC, Biolegend), PDGFRα (APC, Biolegend).
Techniques: Mouse Assay, Staining, Expressing, Flow Cytometry, Cytometry