Methods for the isolation and 3D culture of dermal papilla cells from human hair follicles

Abstract

The dermal papilla is a cluster of mesenchymal cells located at the base of the hair follicle which have a number of important roles in the regulation of hair growth. As a consequence, in vitro models of these cells are widely used to study the molecular mechanisms which underlie hair follicle induction, growth and maintenance. While dermal papilla from rodent hair follicles can be digested prior to cell isolation, the unique extracellular matrix composition found in human dermal papilla renders enzymes such as trypsin and collagenase insufficient for digestion of the dermal papilla into a single cell suspension. As such, to grow human dermal papilla cells in vitro, the papilla has to first be isolated via a micro‐dissection approach from the follicle. In this article we describe the micro‐dissection and culture methods, which we use within our laboratory, for the study of human dermal papilla cells.

Extracellular Vesicles from Activated Dermal Fibroblasts Stimulate Hair Follicle Growth Through Dermal Papilla-Secreted Norrin

“…All human donors provided written informed consent for research. hDPs were microdissected from the bulbar portion of HF under a dissection microscope as previously described . Ten papillae were transferred per well of a 24‐well plate in 500 μl of attachment medium (minimum essential medium (MEM) [1 g/l d ‐glucose] supplemented with 20% fetal bovine serum (FBS), 100 U/ml of penicillin/streptomycin, and 0.5 μg/ml of amphotericin B, all from Gibco, Life Technologies, Courtaboeuf, France).…”

Subpopulations of dermal skin fibroblasts secrete distinct extracellular matrix: implications for using skin substitutes in the clinic

“…Intact dermal papillae were also isolated from the same piece of scalp skin using a microdissection approach to obtain DPfi cultures from the same donors as the Pfi and Rfi . Follicles were transected just above the level of the dermal papilla to isolate end bulbs, which were inverted using 27‐gauge needles to remove the matrix and expose the dermal papilla.…”

Human Derived Immortalized Dermal Papilla Cells With a Constant Expression of Testosterone Receptor

“…Therefore, in vitro culture of DPCs would be useful to find out the molecular target and the screening of pharmaceutical products to treat AGA. DPCs can be prepared from primary cultures of human cells, but sampling and primary cell culture can produce wide variability depending on cell preparation (Topouzi et al, 2017). Furthermore, primary DPCs cannot continuously proliferate because of cellular senescence and the Hayflick limit.…”