Learning Objectives:
Introduction: The epidermal basal stem/progenitor cell maintains homeostasis of epidermis under development, self-renewal and differentiation. In many cases of adult basal stem/progenitor cell regulation, the importance of extracellular signals provided by the surrounding cells are well recognized. Keratinocyte growth factor (KGF) is a mesenchymal-cell-derived paracrine growth factor that specifically participate in tissue development as well as wound repair. In this study, we investigated the effects of over-expressed KGF during epithelial cell proliferation and differentiation by using a cell labeling system.
Methods: After anesthetized ICR mouse Flag-hKGF cDNA driven by a CMV14 promoter was transfected into ear skin with electroporation. The ears with empty vector transfection were used as controls. 5-bromo-2′-deoxyuridine (BrdU) and 5-ethynyl-2′ deoxyuridine (EdU) were administered at different time points before or after KGF expression vector transfection to identify stem cells or progenitor cells, which are believed to divide slowly or to segregate chromosomes asymmetrically. At 1, 4 and 7 days after vector transfection, 3 mice at each time-point were sacrificed. The paraffin sections were used for H&E and immunohistochemistry for Flag, KGF, BrdU and cytokeratin (CK)14. EdU staining was performed according to the manufacturer's protocol (Life Technologies).
Results: Each plasmid was transfected into the epithelial and subepithelial cells, successfully. After KGF transfection, keratin accumulations were observed 3 of 3 ears at 4 days. BrdU(+)EdU(+) cells (stem/progenitor cell) were detected in the upper layer of thickened CK14 positive epithelium in KGF transfected specimens at 4 days.
Conclusions: These findings indicated that KGF overexpression may possibly increase stem or progenitor cell proliferation and block terminal differentiation, resulting in epithelial hyperplasia and stratification.
