Accelerated Wound Healing by Fibroblasts Differentiated from Human Embryonic Stem Cell-Derived Mesenchymal Stem Cells in a Pressure Ulcer Animal Model

Figure 1

Fibrogenic differentiation of human embryonic stem cell-derived mesenchymal stem cells (hESC-MSCs) upon stimulation with connective tissue growth factor (CTGF). hESC-MSCs were differentiated into fibroblasts by treatment with various concentrations of connective tissue growth factor (CTGF) for 4 weeks. Normal skin fibroblasts (Detroit 551) were also used as a positive control. (a) mRNA levels of fibroblast-related genes in hESC-MSCs after CTGF treatment were determined by the real-time polymerase chain reaction (PCR) (, one-way ANOVA; and ). (b) Collagen (Col)1, Col3, fibronectin (FN), and fibroblast-specific protein- (FSP-) 1 mRNA levels were determined by PCR. (c) FN, FSP-1, Col1, and β-actin protein levels in hESC-MSCs following CTGF treatment were determined by immunoblotting. (d) Masson’s trichrome was used to detect collagen fibers. (e) hESC-MSCs were immunostained to detect collagen I (Col1) following CTGF treatment. 4,6-Diamidino-2-phenylindole (DAPI) was used for nuclear counterstaining. (f) Flow cytometry analysis of hESC-MSCs. After expansion of hESC-MSCs and hESC-MSC-Fbs, cells were trypsinized and stained with specific markers for CD29, CD47, CD73, CD90, CD91, CD105, and CD166.

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