Isolation And Characterization Of Sox2-positive Dermal Papilla Cells

Skin-derived precursor cells(SKPs) are of clinical interests because they have a potential of generating neurons, smooth muscle cells, and adipocytes andmay be considered immunologically privileged.However, the frequency decreases fast during adulthood, and primary sphere generation from adult human skin is more problematic. According to reports, SKPs, which can used in repairing the injured CNS, derive from Sox2+ dermal papilla cells. As Sox2 is expressed in nucleus,to get living Sox2 positive dermal papilla cells,we isolated human dermal papilla cells from hair follicles, instructed a lentivirus reporter vector driven by the Sox2 promoter,cultured the transducted dermal papilla cells in the sphere-formation medium,and identified the cells we isolated.Methods: 1. Isolated dermal papilla cells through microdissection of hair follicles plucked form the volunteers, and characterized them by ALP detection 、 RT-PCR and immunofluorescence. 2. Extacted genomic DNA from the cultured umbilical cord mesenchymal stem cells, cloned the fragment of Sox2 promoter,ligated to the T-vector, transformed into competent cells, spreaded on the LB solid plates, and cultured overnight in 37℃. Confirmed the fragment by PCR、enzyme cleavage and sequencing.To get the desired lentivirus vector,weconstructed the previous one with enzyme cleavage and ligation of Sox2 promoter and luciferase fragment. 3. After lentivirus preparation and cell transduction, digested the transducted dermal papilla cells to single ones, cultured them in sphere-formation medium and picked the e GFP-positive spheres under the fluorescence microscopeConlusions: 1. We isolated human dermal papilla cells successfully. And during the in vitro expansion, they were good.2. The lentivirus reporter vector driven by the Sox2 promoter was constructed with success. 3. We succeeded in isolating living Sox2 positive dermal papilla cells.