Figure 6. EB-derived blister fluids support migration of CXCR1⁺ and CXCR2⁺ ADSC.

(a) Representative FACS profiles of CXCR1⁺ and CXCR2⁺ ADSC in the Transwell migration assay. Percentage of retaining (top chamber) and migrated (bottom chamber) cells is indicated in inserts: Orange - cell migration without BF (Control); Blue and Green - cell migration toward BF at 1:20 and 1:60 dilution, respectively. Histogram markers define a threshold of chemokine-receptor positive cells. (b) Analysis of retaining and migrating CXCR1⁺ and CXCR2⁺ ADSC in the Transwell chambers. Data were collected from 2 independent experiments in triplicates. Data are presented as scatter plots depicting percentage of CXCR1⁺ and CXCR2⁺ cells. Statistical significance (p-value) is shown above the plots. (c) Analysis of BF-induced ADSC migration in collagenous matrix. Top left - scheme of the in-matrix migration unit. PM - porous membrane; ADSC - ADSC-containing collagenous gel; Col - separating collagenous layer; BF - blister fluid-containing collagenous gel; CM - culture media. Arrow indicates direction of migration. Representative images of cross sections of migration units are shown on the right: top row - migration of the unselected ADSC toward culture media (CM) and blister fluids (BF); bottom row - migration of CXCR1⁺ and CXCR2⁺ ADSC toward BF. Blue - DAPI nuclear staining; Green - vibrant DiO-labeled ADSC; Red - CXCR1⁺ or CXCR2⁺ ADSC detected by indirect immunofluorescence. Scale bar - 100 μm. Single channel images are presented in Supplementary materials (Fig. S6c). Bottom left - quantitative assessment of ADSC migration in collagenous matrix. Migration is calculated as percentage of total (open columns) and CXCR1- or CXCR2-positive (red columns) ADSC migrated less than 100 or more than 200 μm per microscopic field ± SD. Conditions are indicated below the columns.