The dermal papilla (DP) is a cluster of mesenchymal cells located at the bottom of the hair follicle. Cells within the DP interact with numerous other cell types within the follicle, including epithelial stem cells, matrix cells, and melanocytes, regulating their function. The diameter of the DP is directly proportional to the width of the hair shaft, and a decrease in both cell number and DP size is observed in hair loss conditions such as androgenetic alopecia. Conversely, microdissected ex vivo DP can instruct growth of de novo hair follicles. The study of DP cells and their role in human hair growth is often hampered by the technical challenge of DP isolation and culture. Here we describe a method used within our research group for isolating DP from human hair follicles.