Identification and Sorting of Epidermal Progenitors

Each sample was transferred to a collection tube and resuspended in PBS/1% BSA/5mM EDTA. Fluorescently tagged antibodies were added at concentrations presented in Table 3 and incubated for 30 min on ice. Labeled cells were washed three times with PBS to remove unbound antibody and resuspended in flow buffer (PBS with 2% BSA, 1 mM EDTA, and 1 μg/mL DAPI). Cells were sorted on a BD FacsAria III. LEps were defined as CD133+ /CD271− cells, and MEps were defined as CD133−/CD271+ cells, with DAPI + cells discarded. Compared to cells from standard 2-D cultures, postconfluent cells exhibit marked autofluorescence.