bs-4836r  (Bioss)

Journal: Stem Cell Research & Therapy

Article Title: Chemotaxis-driven disease-site targeting of therapeutic adult stem cells in dystrophic epidermolysis bullosa

doi: 10.1186/s13287-016-0388-y

Figure Lengend Snippet: Analysis of wild-type and DEB-affected skin infiltration with leukocytes. a H&E staining and indirect immunofluorescent analysis of skin infiltration with leukocytes. Detected antigens are shown in respective colors above the panels (CD45, green ; CXCR2, green ; CD11b, red ; colocalization of CD45 or CXCR2 and CD11b, yellow ). Nuclei were counterstained with DAPI ( blue ). bc blister cavity. Scale bar = 100 μm. b Quantitation of the skin-infiltrating cells. Data presented as an average number or percentage of cells (as indicated on the Y axis) per microscopic field ± SD. * p <0.05. ** p > 0.05. Data collected from at least 15 independent microscopic fields. CD-positive cell types are shown above the columns. H&E hematoxylin and eosin, WT wild-type skin, NB nonblistering, DEB dystrophic epidermolysis bullosa, Bl blistering DEB-affected skin

Article Snippet: For indirect immunofluorescent analysis, cross-sections were stained with the following primary antibodies: goat anti-mouse CD45 polyclonal (BD Bioscience), rat anti-mouse CD11b monoclonal (BD Bioscience), rabbit anti-CXCR2 polyclonal (Bioss, Woburn, MA, USA), mouse anti-FLuc monoclonal (Abcam, Cambridge, MA, USA), rabbit anti-type VII collagen polyclonal (Millipore, Billerica, MA, USA), and goat anti-type IV collagen polyclonal (Millipore).

Techniques: Staining, Quantitation Assay

Journal: Stem Cell Research & Therapy

Article Title: Chemotaxis-driven disease-site targeting of therapeutic adult stem cells in dystrophic epidermolysis bullosa

doi: 10.1186/s13287-016-0388-y

Figure Lengend Snippet: Systemic transplantation of genetically engineered CXCR2 + -mADSC into DEB-affected newborn mice. a Characterization of CXCR2 expression in primary mADSC and cells engineered to overexpress FLuc (FLuc + -mADSC) and FLuc and Cxcr2 genes (FLuc + CXCR2 + -mADSC), respectively, by FACS and indirect immunofluorescent detection. Analyses were performed using gene-specific antibodies. (Fluc, red ; CXCR2, green ; colocalization of FLuc and CXCR2, yellow ; nuclei were counterstained with DAPI). b In-vivo imaging of systemically administered FLuc + CXCR2 + -mADSC and native FLuc + -mADSC into newborn Col7a1 –/– and wild-type mice, respectively, 24 and 48 hours after transplantation (as indicated on the left ). White and orange arrows point to the blister location. Type of transplanted cells indicated below the image of a representative mouse. c Representative micrographs of H&E-stained sections of the skin collected from blistering sites (ventral skin and paws) of FLuc + -mADSC and FLuc + CXCR2 + -mADSC transplanted animals, respectively. Black arrowheads point to infiltrating cells. bc blister cavity. d Indirect immunofluorescent analysis of mADSC recruitment to blistering sites (indicated to the left of the panels). Type of mADSC used for transplantation indicated above the panels. Detected antigens, FLuc, CXCR2, and Col7, are shown below the panels in corresponding colors. Scale bar = 100 μm. bc blister cavity, white arrows point to the stretches of the BMZ-associated type VI collagen. e Quantitation of cell recruitment to blistering sites. Y axis represents the percentage of positive cells per microscopic field. The FLuc + , CXCR2 + , and Col7 + cells are shown below the columns. Data shown as mean ± SD. * p <0.05. ** p > 0.05. DEB dystrophic epidermolysis bullosa, mADSC mouse adipose-derived stem cells

Article Snippet: For indirect immunofluorescent analysis, cross-sections were stained with the following primary antibodies: goat anti-mouse CD45 polyclonal (BD Bioscience), rat anti-mouse CD11b monoclonal (BD Bioscience), rabbit anti-CXCR2 polyclonal (Bioss, Woburn, MA, USA), mouse anti-FLuc monoclonal (Abcam, Cambridge, MA, USA), rabbit anti-type VII collagen polyclonal (Millipore, Billerica, MA, USA), and goat anti-type IV collagen polyclonal (Millipore).

Techniques: Transplantation Assay, Expressing, In Vivo Imaging, Staining, Quantitation Assay, Derivative Assay

Journal: Stem Cell Research & Therapy

Article Title: Chemotaxis-driven disease-site targeting of therapeutic adult stem cells in dystrophic epidermolysis bullosa

doi: 10.1186/s13287-016-0388-y

Figure Lengend Snippet: Transplantation of CXCR2-enriched mADSC into DEB-affected mice. a In-vivo live imaging of neonatal DEB mice and 2 and 7 days after IP transplantation of the DiI-labeled native and CXCR2 + -mADSC ( red ), respectively. b Direct fluorescent detection of the DiI-mADSC in the blistering skin of the recipient mice at 7 days after transplantation. Blister cavity ( bc ) outlined by dotted line . c Indirect immunofluorescent detection of type VII and type IV collagens in the skin of mADSC-treated and control mice (indicated above the panels). Detected antigens (in corresponding color) are shown to the left of the panels. Upper row , monochrome images of the sections stained with anti-type VII collagen antibodies. White arrowheads point to the BMZ. Blue , DAPI nuclear staining. Scale bar = 100 μm. DEB dystrophic epidermolysis bullosa, mADSC mouse adipose-derived stem cells

Article Snippet: For indirect immunofluorescent analysis, cross-sections were stained with the following primary antibodies: goat anti-mouse CD45 polyclonal (BD Bioscience), rat anti-mouse CD11b monoclonal (BD Bioscience), rabbit anti-CXCR2 polyclonal (Bioss, Woburn, MA, USA), mouse anti-FLuc monoclonal (Abcam, Cambridge, MA, USA), rabbit anti-type VII collagen polyclonal (Millipore, Billerica, MA, USA), and goat anti-type IV collagen polyclonal (Millipore).

Techniques: Transplantation Assay, In Vivo, Imaging, Labeling, Staining, Derivative Assay