Controllable production of transplantable adult human high-passage dermal papilla spheroids using 3D matrigel culture.

We have succeeded in culturing human dermal papilla (DP) cell spheroids and developed a three-dimensional (3D) Matrigel (basement membrane matrix) culture technique that can enhance and restore DP cells unique characteristics in vitro. When 1 × 10(4) DP cells were cultured on the 96-well plates precoated with Matrigel for 5 days, both passage 2 and passage 8 DP cells formed spheroidal microtissues with a diameter of 150-250 µm in an aggregative and proliferative manner. We transferred and recultured these DP spheroids onto commercial plates. Cells within DP spheres could disaggregate and migrate out, which was similar to primary DP. Moreover, we examined the expression of several genes and proteins associated with hair follicle inductivity of DP cells, such as NCAM, Versican, and α-smooth muscle actin, and confirmed that their expression level was elevated in the spheres compared with the dissociated DP cells. To examine the hair-inducing ability of DP spheres, hair germinal matrix cells (HGMCs) and DP spheres were mixed and cultured on Matrigel. Unlike the dissociated DP cells and HGMCs cocultured in two dimensions, HGMCs can differentiate into hair-like fibers under the induction of the DP spheres made from the high-passage cells (passage 8) in vitro. We are the first to show that passage 3 human HGMCs differentiate into hair-like fibers in the presence of human DP spheroids. These results suggest that the 3D Matrigel culture technique is an ideal culture model for forming DP spheroids and that sphere formation partially models the intact DP, resulting in hair induction, even by high-passage DP cells.

Promotional effect of platelet-rich plasma on hair follicle reconstitution in vivo.

BACKGROUND: Platelet-rich plasma (PRP) containing various growth factors has attracted attention in various medical fields. PRP has recently been used during hair transplantation to increase hair density. OBJECTIVE: To investigate the effects of PRP on hair follicle (HF) reconstitution. METHODS AND MATERIALS: Freshly isolated epidermal cells and cultured dermal papilla cells (DPCs) were mixed with various concentrations of activated PRP and transferred to a grafting chamber that was implanted onto the dorsal skin of nude mice. The chambers were removed 1 week after grafting, and HF formation was monitored for 4 weeks. RESULTS: We observed a significant difference (p < .05) in the number of newly formed follicles in the area of reconstituted skin (344 ± 27 with 10% PRP vs 288 ± 35 without PRP). PRP also shortened the time of hair formation significantly; the first hairs were observed in 18 ± 1 days using 10% PRP, versus 20 ± 1 days without PRP. CONCLUSION: A considerable effect of PRP on the time of hair formation and the yield of HF reconstitution was observed in this study. Considering the limited evidence available to judge its efficacy, further studies are required to investigate the mechanism of action of PRP.

[Effect of PRP on the proliferation of dermal papilla cells and hair follicle regeneration in mice].

OBJECTIVE: To investigate the effects of platelet-rich plasma (PRP) on the proliferation of dermal papilla cells (DPCs) and hair follicle regeneration. METHODS: PRP was prepared using the double-spin method and applied to DPCs. The proliferative effect of activated PRP on DPCs was measured using MTT assay. To understand the influence of activated PRP on the hair-inductive capacity of DPCs, freshly isolated epidermal cells and DPCs of passage 4 were resuspended, mixed with various concentrations of a PRP (0%, 5% or 10%) and were then transferred to a grafting chamber, which was implanted onto the dorsal skin of nude mice. The chambers were removed 1 week after grafting and HF formation was monitored for 4 weeks; the graft site was harvested and processed for histological examination. RESULTS: Activated PRP increased the proliferation benefited the aggregative growth of DPCs. There are significant difference in the yield of hair follicles compared with 10% PRP (344 +/- 27) with 0% PRP (288 +/- 35) in the area of reconstituted skin (P < 0.05). The areas treated with PRP demonstrated an increase in hair follicles density of 19.4%. Ten percent PRP (18 +/- 1) d also can significantly shorten the time of hair formation, compared with 0% PRP (20 +/- 1) d (P < 0.05). CONCLUSIONS: There is a considerable effect of PRP on the time of hair formation and the yield of hair follicles reconstitution.

[Inhibition effect of 6-gingerol on hair growth].

OBJECTIVE: To investigate the effect of 6-gingerol, the main active component of ginger, on hair shaft elongation in vitro and hair growth in vivo. METHODS: Firstly, Hair follicles were co-cultured with 3 different concentration of 6-gingerol for 5 days and hair elongation in three groups was measured. Secondly, The proliferative effect of 6-gingerol on DPCs was measured using MTT assay. Thirdly, the expression of Bcl-2 and Bax in DPCs were measured using Western blotting. In vivo study, the influence of 6-gingerol on hair growth in C57BL/6 rats was measured through topical application of 6-gingerol on the dorsal skin of each animal. RESULTS: The length of hair shaft in 20 microg/ml 6-Gingerol group (0.50 +/- 0.08 mm) is less than 0 microg/ml (0.66 +/- 0.19) mm and 10 microg/ml (0.64 +/- 0.03) mm 6-Gingerol group (P < 0.05). In cell culture, compared to 0 microg/ml and 5 microg/ml 6-Gingerol, 10 microg/ml 6-Gingerol can significantly inhibited the proliferation of DPCs (P < 0.05). Along with the growth inhibition of DPCs by 6-gingerol, the Bax/Bcl-2 ratio increased obviously. In vivo study, the hair length and density decreased a lot after using 1 mg/ml 6-gingerol. CONCLUSIONS: 6-Gingerol can suppress human hair shaft elongation because it has pro-apoptotic effects on DPCs via increasing Bax/Bcl-2 ratio. It might inhibit hair growth by prolonging the telogen stage in vivo.