ITRAQ-Based Quantitative Proteomic Comparison of Early- and Late-Passage Human Dermal Papilla Cell Secretome in Relation to Inducing Hair Follicle Regeneration.

Huan Zhang,Chang-Min Lin,Ning-Xia Zhu,Yang Liu,Bo-Zhi Cai,Yang Zeng,Keng Huang,Yan-Ping Yuan,Yan-Ming Xu,Jan Peter Tuckermann

doi:10.1371/journal.pone.0167474

Abstract

Alopecia is an exceedingly prevalent problem that lacks effective therapy. Recently, research has focused on early-passage dermal papilla cells (DPCs), which have hair inducing activity both in vivo and in vitro. Our previous study indicated that factors secreted from early-passage DPCs contribute to hair follicle (HF) regeneration. To identify which factors are responsible for HF regeneration and why late-passage DPCs lose this potential, we collected 48-h-culture medium (CM) from both of passage 3 and 9 DPCs, and subcutaneously injected the DPC-CM into NU/NU mice. Passage 3 DPC-CM induced HF regeneration, based on the emergence of a white hair coat, but passage 9 DPC-CM did not. In order to identify the key factors responsible for hair induction, CM from passage 3 and 9 DPCs was analyzed by iTRAQ-based quantitative proteomic technology. We identified 1360 proteins, of which 213 proteins were differentially expressed between CM from early-passage vs. late-passage DPCs, including SDF1, MMP3, biglycan and LTBP1. Further analysis indicated that the differentially-expressed proteins regulated the Wnt, TGF-β and BMP signaling pathways, which directly and indirectly participate in HF morphogenesis and regeneration. Subsequently, we selected 19 proteins for further verification by multiple reaction monitoring (MRM) between the two types of CM. These results indicate DPC-secreted proteins play important roles in HF regeneration, with SDF1, MMP3, biglycan, and LTBP1 being potential key inductive factors secreted by dermal papilla cells in the regeneration of hair follicles.

Highlights

Alopecia is an exceedingly prevalent problem that affects psychological well-being, and endangers certain inherent functions of the skin [1]
Cell-based hair regeneration has been investigated as a possible alternative, and various experimental assays have shown that early-passage dermal papilla cells (DPCs) can induce hair follicle (HF) regeneration both in vitro and in vivo [2]
In order to determine whether DPC-secreted proteins induce HF regeneration, we collected passage 3 and 9 DPC-culture medium (CM), concentrated the CM through an Amicon filter, and subcutaneously injected the concentrated (2 mg/ml) CM into NU/NU mice

Summary

Introduction

Alopecia is an exceedingly prevalent problem that affects psychological well-being, and endangers certain inherent functions of the skin [1]. The dermal papilla (DP), formed at the base of the hair follicle, is a unique tissue surrounded by epithelial matrix cells and is essential in the control of hair growth, formation and cycling. The tendency to aggregate is one of the significant characteristics of DPCs and is associated with biological function. This ability to aggregate and capacity to induce HF differentiation is lost after several passages of culture [2]. The molecular mechanisms of sustaining the inductive ability of DPCs have not been explored completely, the ability of conditioned medium to sustain the formation of aggregates implies that secreted proteins, from cells into the culture medium, may play roles in preserving the inductive ability of DPC

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