Journal: Cell stem cell

Article Title: Gli1+ mesenchymal stromal cells are a key driver of bone marrow fibrosis and an important cellular therapeutic target

doi: 10.1016/j.stem.2017.03.008

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

Journal: International Journal of Stem Cells

Article Title: CD73, CD90, CD105 and Cadherin-11 RT-PCR Screening for Mesenchymal Stem Cells from Cryopreserved Human Cord Tissue

doi: 10.15283/ijsc17015

Figure Lengend Snippet: MSC expansion from cryopreserved UCT expresses CD73, CD90 and CD105 cell surface markers. MSC expanded from cryopreserved UCT sections were harvested and screened by flow cytometry for (A) positive CD90, CD73, CD105 and CD44 and checked against (B) negative isotype CD90, CD73, CD105 and CD44. (C) Combined positive CD90, CD73, CD105 and CD44 population for MSC expansion from fresh and frozen are presented as percent mean±SD from n=6.

Article Snippet: Filtered cells were dispensed to 100 μ l aliquots, incubated with mesenchymal stem cell marker CD73, CD90 and CD105 (Stemflow, Becton Dickinson) and shielded from light for 20 min at room temperature.

Techniques: Flow Cytometry, Cytometry

Journal: International Journal of Stem Cells

Article Title: CD73, CD90, CD105 and Cadherin-11 RT-PCR Screening for Mesenchymal Stem Cells from Cryopreserved Human Cord Tissue

doi: 10.15283/ijsc17015

Figure Lengend Snippet: MSC expansion from fresh and cryopreserved UCT express CD90, CD73 and CD105 mRNA. MSC expanded from (A) fresh and (B) cryopreserved UCT from 2 different donors were processed for RNA, converted to cDNA and screened for CD73, CD90 and CD105 by RT-PCR. GAPDH was used as an internal control and data are expressed as mean fold expression against GAPDH±SD from 3 experiments. Representative reactions were resolved on a 1.5% agarose gel and captured on an imager.

Article Snippet: Filtered cells were dispensed to 100 μ l aliquots, incubated with mesenchymal stem cell marker CD73, CD90 and CD105 (Stemflow, Becton Dickinson) and shielded from light for 20 min at room temperature.

Techniques: Reverse Transcription Polymerase Chain Reaction, Expressing, Agarose Gel Electrophoresis

Journal: International Journal of Stem Cells

Article Title: CD73, CD90, CD105 and Cadherin-11 RT-PCR Screening for Mesenchymal Stem Cells from Cryopreserved Human Cord Tissue

doi: 10.15283/ijsc17015

Figure Lengend Snippet: Fresh and Cryopreserved UCT expresses CD73, CD90 and CD105 mRNA. (A) Fresh and (B) cryopreserved UCT from 4 different donors were processed for RNA, converted to cDNA and screened for CD73, CD90 and CD105 by RT-PCR. GAPDH was used as an internal control and data are expressed as mean fold expression against GAPDH±SD from 3 experiments. Representative reactions were resolved on a 1.5% agarose gel and captured on an imager.

Article Snippet: Filtered cells were dispensed to 100 μ l aliquots, incubated with mesenchymal stem cell marker CD73, CD90 and CD105 (Stemflow, Becton Dickinson) and shielded from light for 20 min at room temperature.

Techniques: Reverse Transcription Polymerase Chain Reaction, Expressing, Agarose Gel Electrophoresis