Optimal stimulation toward the dermal papilla lineage can be promoted by combined use of osteogenic and adipogenic inducers

Abstract

Dermal papilla cells (DPCs) play crucial roles in hair regeneration, but they readily lose their hair‐forming ability during in vitro culture. Although the formation of spheroids partially restores the ability, shrinkage of the spheroids makes it difficult to maintain cellular viability. To address this problem, we stimulated DPCs with factors known to induce adipogenic and/or osteogenic differentiation, because DPCs share unique gene expression profiles with adipocytes and osteocytes. We isolated DPCs from versican (vcan)–GFP mice, in which GFP is expressed under the control of a vcan promoter, which is strongly active in DPCs of anagen hair follicles. GFP fluorescence was most intense when the spheroids were made from DPCs cultured in a half‐diluted combination of adipogenic and osteogenic media (CAO1/2), a Dulbecco’s modified Eagle’s medium‐based medium that contains 10% FBS, 275 nm dexamethasone, 2.5 mm β‐glycerol phosphate, 12.5 µg·mL−1 ascorbic acid, 0.125 µm isobutylmethylxanthine and 2.5 ng·mL−1 insulin. The dose of each additive used was less than the optimal dose for adipogenic or osteogenic differentiation, and shrinkage of the spheroids was avoided through the addition of fibroblast growth factor 2 and platelet‐derived growth factor‐AA to CAO1/2. In addition, the gene and protein expression of vcan, osteopontin, alkaline phosphatase and α‐smooth muscle actin in the spheroids were augmented to levels similar to those of the intact dermal papillae, which exhibited restored hair‐forming activity. In conclusion, a combination of certain adipogenic and osteogenic inducers, together with fibroblast growth factor 2 and platelet‐derived growth factor‐AA, can promote differentiation toward the DPC lineage.

Dermal Papilla Cell-Derived Extracellular Vesicles Increase Hair Inductive Gene Expression in Adipose Stem Cells via β-Catenin Activation

“…We previously reported that CAO1/2FP medium (a combination of adipogenic and osteogenic inducers at low concentrations, with FGF-2 and PDGF-AA) was adequate for DPC in vitro culture and prevented DPC shrinkage by maintaining extracellular matrix components, versican, and other hair induction-related gene expression [ 22 ]. We used CAO1/2FP in the current study as a differentiation medium for ASCs.…”

“…CAO1/2FP seemed to be appropriate for ASCs and DPCs to transform ASCs into DPC-like cells. DPCs express characteristic genes, such as vcan , sma , sox2 , ctnb , Alpl , and so on, for HF morphogenesis into the postnatal hair cycle in mammals [ 22 , 30 , 31 ]. These genes are also expressed in non-DPCs, such as dermal sheath cells and dermal fibroblasts; however, combinations of these molecules are cell-type specific.…”

“…CAO1/2FP was prepared as previously reported [ 22 ]. Briefly, Dulbecco’s modified Eagle’s medium (DMEM) with 10% FBS (DMEM-FBS) adipogenic and osteogenic media were mixed in a 1:1 ratio and addressed as a combination of adipogenic and osteogenic media (CAO).…”

Versican and versikine: The dynamism of the extracellular matrix

“…64,65 Dermal papillae cells isolated from Vcan promoter driven-GFP mice were applied to a spheroid culture, and a half-diluted combination of adipogenic and osteogenic medium together with FGF2 and PDGF-AA promoted expression of dermal papillae-specific gene expression. 66 Although several researchers used Vcan expression as a dermal papillae cell marker, 67 only a limited number of reports have implied its role in dermal papillae formation. The Vcan Δ3/Δ3 dermal papillae show decreased levels of Vcan deposition as well as the number of hair follicles.…”

“…In contrast, it is absent in vellus hair follicles of bald male patterns even in anagen 64,65 . Dermal papillae cells isolated from Vcan promoter driven‐GFP mice were applied to a spheroid culture, and a half‐diluted combination of adipogenic and osteogenic medium together with FGF2 and PDGF‐AA promoted expression of dermal papillae‐specific gene expression 66 . Although several researchers used Vcan expression as a dermal papillae cell marker, 67 only a limited number of reports have implied its role in dermal papillae formation.…”