Figure 1

Characterization of stem cell populations obtained from skin biopsies or already established skin fibroblast cultures. (A-D) Phase contrast images of hSKIN-MASC at the third passage in culture: A-B) hSKIN-MASC obtained from skin biopsies of a healthy donor (A) and a NPC patient (B). C-D) hSKIN-MASC obtained from already established skin fibroblasts from healthy donor (C) and NPC patient (D). (E) Surface immunophenotype: representative flow cytometry histograms of skin derived stem cell cultures. Plots show isotype control IgG-staining profile (green histograms) versus specific antibody staining (red histograms). (F-H) Pluripotent state specific transcription factor expression: representative fluorescence images of Oct-4 (green fluorescence; F), Nanog (green fluorescence; G) and Sox-2 expression (green fluorescence; H) localized in the nuclei of skin derived stem cell cultures. Nuclei are depicted by the blue fluorescence of DAPI staining (F-H). I) Quantification of the percentage of cells expressing Oct4 (white bars), Nanog (black bars) and Sox-2 (gray bars) in hSKIN-MASC obtained both from healthy donors (CTRL, n = 3) and NPC (n = 3) skin biopsies or healthy donors (CTRL, n = 3) and NPC (n = 3) skin fibroblast cultures. At least 400 cells have been counted for each cell line. Data are presented as mean ± SD of 3 independent experiments. (J-K) Nestin expression (red fluorescence) in hSKIN-MASC obtained from a healthy donor (J) and a NPC patient (K). (L-N) Unesterified cholesterol accumulation: images obtained after performing the filipin staining (blue fluorescence) in hSKIN-MASC derived from a healthy donor skin biopsy (L), a NPC patient skin biopsy (M) and a NPC skin fibroblast cell line (N), respectively.