The rapid development of biomimetic materials in the field of regenerative medicine has made it possible to mimic natural cellular environments and allow in vitro systems to replace in vivo systems. In particular, the emergence of hexanoyl glycol chitosan (HGC) is playing an important role the development of 3-dimensional (3D) cell culture and tissue engineering. We employed HGC-coated dishes to cultivate human hair follicle dermal papilla (HDP) cells in 3D spheroids, assessing their ability to form hair-like structures. The study also tested the effect of minoxidil on these spheroids and explored their integration into artificial skin models. HDP cell spheroids successfully formed hair-like structures within the 3D culture. Minoxidil treatment showed enhanced hair growth in spheroids cocultured with keratinocytes. In addition, transplantation of these spheroids into artificial skin led to the formation of functional papilla structures, suggesting a closer mimicry of human skin. Hair-like structure formation and successful integration into artificial skin models pave the way for innovative approaches in hair loss treatment research, cosmetic, and pharmaceutical evaluations and skin restoration therapies.