Fig. 3

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