human fibronectin matrix  (Developmental Studies Hybridoma Bank)

Journal: Journal of biomedical materials research. Part A

Article Title: Development of hybrid scaffolds with natural extracellular matrix deposited within synthetic polymeric fibers 3

doi: 10.1002/jbm.a.36078

Figure Lengend Snippet: PDTEC:PEG polymer fiber mat were washed in dH2O to remove PEG. Mouse NIH 3T3 fibroblasts were grown for 7 days within the PDTEC fiber mat. Cells were fixed and stained with R457 anti-fibronectin antiserum (green) and DAPI to stain the nuclei (blue). Confocal images shown are (A) maximum intensity projection (scale bar = 50 μm), (B) maximum intensity projection of the view orthogonal to (A) showing the depth of the matrix within the scaffold, and (C) an alpha-blended volume view (scale bar = 25 μm).

Article Snippet: After 24 h, cultures were fixed, permeabilized for 15 min in 0.5% Triton X (Sigma) in PBS, and stained to visualize the decellularized matrix with R457 polyclonal antiserum diluted 1:100, actin cytoskeleton with either rhodamine-phalloidin or Texas Red-X phalloidin (ThermoFisher) 1:50, nuclei with 1 μg/mL DAPI, or newly deposited human fibronectin matrix with HFN7.1 monoclonal anti-fibronectin antibody (DSHB, University of Iowa) diluted at 1:100.

Techniques: Polymer, Staining

Journal: Journal of biomedical materials research. Part A

Article Title: Development of hybrid scaffolds with natural extracellular matrix deposited within synthetic polymeric fibers 3

doi: 10.1002/jbm.a.36078

Figure Lengend Snippet: After PEG removal, NIH3T3 cells were cultured within the scaffolds for 7 d and then fixed and stained with anti-fibronectin antibodies (green) and DAPI (blue). A and C are the same microscope field; (A) shows the bright field image of the synthetic fibers and (C) shows the fluorescence signal of matrix and nuclei. In parallel, a similar culture was decellularized and then fixed and stained as above. B and D show the same field by bright field and fluorescence imaging. The fiber mat organization is not affected by the decellularization procedure (compare A and B). The absence of DAPI staining in D shows that cells are removed by decellularization while matrix fibrils remain in place. Scale bars = 50 μm.

Article Snippet: After 24 h, cultures were fixed, permeabilized for 15 min in 0.5% Triton X (Sigma) in PBS, and stained to visualize the decellularized matrix with R457 polyclonal antiserum diluted 1:100, actin cytoskeleton with either rhodamine-phalloidin or Texas Red-X phalloidin (ThermoFisher) 1:50, nuclei with 1 μg/mL DAPI, or newly deposited human fibronectin matrix with HFN7.1 monoclonal anti-fibronectin antibody (DSHB, University of Iowa) diluted at 1:100.

Techniques: Cell Culture, Staining, Microscopy, Fluorescence, Imaging

Journal: Journal of biomedical materials research. Part A

Article Title: Development of hybrid scaffolds with natural extracellular matrix deposited within synthetic polymeric fibers 3

doi: 10.1002/jbm.a.36078

Figure Lengend Snippet: Human MSCs were plated on a hybrid ECM – polymer scaffold or on a polymer scaffold without ECM. Cells were allowed to spread for 24 h, then cell shapes were visualized by staining actin filaments with rhodamine-phalloidin (red) and DAPI (blue). Samples were also stained with anti-fibronectin antiserum to show the NIH 3T3 matrix within the scaffold. B, C, E, F are higher magnification images of the samples in A and D. Scale bars = 50 μm.

Article Snippet: After 24 h, cultures were fixed, permeabilized for 15 min in 0.5% Triton X (Sigma) in PBS, and stained to visualize the decellularized matrix with R457 polyclonal antiserum diluted 1:100, actin cytoskeleton with either rhodamine-phalloidin or Texas Red-X phalloidin (ThermoFisher) 1:50, nuclei with 1 μg/mL DAPI, or newly deposited human fibronectin matrix with HFN7.1 monoclonal anti-fibronectin antibody (DSHB, University of Iowa) diluted at 1:100.

Techniques: Polymer, Staining

Journal: Journal of biomedical materials research. Part A

Article Title: Development of hybrid scaffolds with natural extracellular matrix deposited within synthetic polymeric fibers 3

doi: 10.1002/jbm.a.36078

Figure Lengend Snippet: Cells were plated as in Figure 5 but then grown for 7 d before staining with rhodamine-phalloidin and DAPI. Human MSC-produced and assembled fibronectin was detected with a human fibronectin-specific monoclonal antibody. Considerably more fibronectin matrix was deposited by cells attached to the hybrid ECM scaffold than to the scaffold alone (compare A and B with D and E). Note that the fibers remain intact throughout the 7 d culture (C, F). Scale bar = 50 μm.

Article Snippet: After 24 h, cultures were fixed, permeabilized for 15 min in 0.5% Triton X (Sigma) in PBS, and stained to visualize the decellularized matrix with R457 polyclonal antiserum diluted 1:100, actin cytoskeleton with either rhodamine-phalloidin or Texas Red-X phalloidin (ThermoFisher) 1:50, nuclei with 1 μg/mL DAPI, or newly deposited human fibronectin matrix with HFN7.1 monoclonal anti-fibronectin antibody (DSHB, University of Iowa) diluted at 1:100.

Techniques: Staining, Produced