Isolation of fresh pelage dermal papilla from versican–GFP transgenic mouse. (a) Fluorescence microscopy of frozen section from newborn versican–GFP transgenic mouse skin. Line indicates hair follicle outline. (b) Partially dissociated dermal cell suspensions. GFP-positive DP at the bottom of undissociated follicles. (c) Complete dissociated cell suspension. DP-derived cells are now seen as single cells. (d) Undissociated follicle pellet removed from cell suspension. No DP associated with the remaining preformed follicle, indicating most of the DP cells were released into the cell suspension. (e–f) Cell-sorting chart of the dermal cells from versican–GFP transgenic mouse skin (e) and postsorting analysis of sorted GFP-positive cell pools (f). Upper windows indicate cells collected as GFP-positive and lower windows indicate those collected as GFP-negative cells. (g) RT-PCR analysis of sorted cells and subsequent culture mRNA expression. −, Negative sorted fresh cells; +, positive sorted fresh cells; +P1, +P2, +P3, passaged culture from GFP-positive sorted cells. Ver, mouse versican primers. (h and i) Cultured DP from positive sorted cells. (h) Primary culture day 1. (i) Primary culture day 4. (b–d, h, and i) Merged images from fluorescence and phase-contrast microscopy.