Figure 5.

Further analysis of derived keratinocytes by global gene expression and transplantation. A. Cluster of global gene expression profiles during keratinocyte differentiation. RNA samples are collected at each stage of keratinocyte differentiation, and gene expression was analyzed by microarray. The global gene expression is clustered by Complete-linkage clustering method. B. Expression of epidermal-specific genes through the differentiation process. Epidermal specific genes were selected according to published dataset, and the pattern of gene expression was displayed by stage of differentiation. C. hESC-derived keratinocyte (hKC) transplantation in mouse excisional wound model. a. Representative image of mouse excisional wound and sites of hKC application. b. Representative image of the wound right after transplantation. Exposed wound areas were marked with yellow dash line. c. Representative image of the wound seven days after wounding and cell transplantation. The skin area was processed for further study. d. H&E staining of the wound healing site seven days after keratinocyte transplantation. Square marks the area shown as high magnification image on the right. Scale bar, 400µm on the left; 100µm on the right. D. Immunostaining of KRT14, KRT1, KRT10, IVL and FLG (red fluorescence) on serial skin sections. Samples were collected seven days after transplantation of keratinocytes derived from GFP-labeled hESCs (green fluorescence). Blue fluorescence, nuclear staining with Hoechst. Scale bar, 100µm on the left; 30µm on the right. E. The percentage of GFP-positive cells expressing specific maturation marker genes, representing keratinocyte maturation in vivo. n=6. Data are presented as the mean±SD of three independent experiments.