Human Umbilical Cord Wharton Jelly-Derived Adult Mesenchymal Stem Cells, in Biohybrid Scaffolds, for Experimental Skin Regeneration

Figure 1

Construction and in vitro characterization of the dermal equivalent. (a) hUCMS morphological features in vitro. (b) Schematic representation of methods to constitute DE, with phase-contrast representative images of the scaffold (S) and DE. (c) The viability test, on the scaffold and DE at the indicated time in comparison with the cell-deprived fibrin scaffold, shows stable values at two different times. (d) Representative SEM images to illustrate the intimate connection between cell and fibrin and the 3D porous structure of fibrin itself. (e) Upper: representative histological H&E stain of the entire DE. Lower: magnification pictures of H&E staining, Masson staining, and keratin staining (left to right). (f) The representative TEM image to illustrate the superficial cell layer seeded on the fibrin scaffold; two cells are well identifiable and it is possible to see their cell membrane very closely. (g) Immunohistochemistry examination of DE in comparison with that of hUCMS maintained in culture conditions for the indicated markers. Ki67 positivity percentages in relation to superior cell multilayer or inferior entrapped cells, in comparison with the cell maintained in normal culture conditions. There is an evident decrease in Ki67-positive cells after DE construction (). (h) qPCR analysis for the indicated markers of DE in comparison with hUCMS alone was used as a calibrator. The expression of each marker, relative to its own control, is equal to 1. HPRT1 served for the control. All results appear as mean ± SD of three independent experiments ().

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