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

PLoS One. 2016 Dec 1;11(12):e0167474. doi: 10.1371/journal.pone.0167474. eCollection 2016.


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.

MeSH terms

  • Animals
  • Biglycan / metabolism
  • Chemokine CXCL12 / metabolism
  • Computational Biology / methods
  • Culture Media, Conditioned / metabolism
  • Dermis / cytology*
  • Dermis / metabolism*
  • Hair Follicle / physiology*
  • Humans
  • Latent TGF-beta Binding Proteins / metabolism
  • Matrix Metalloproteinase 3 / metabolism
  • Mice
  • Proteome*
  • Proteomics* / methods
  • Regeneration*
  • Reproducibility of Results


  • Biglycan
  • Chemokine CXCL12
  • Culture Media, Conditioned
  • Latent TGF-beta Binding Proteins
  • Proteome
  • Matrix Metalloproteinase 3

Grant support

This work was supported by grants from the National Science Foundation of China ( (Nos. 81372084 to KH, 81171832 to CML and 81501685 to NXZ), the Guangdong Province Outstanding Young Teacher Training Program ( (No. Yq2013078 to CML), the Guangdong Scientific Research Funding Program ( (No. 2014A020211024 to CML), and by the National Science Foundation of Guangdong Province ( (No.2016A030310078 to NXZ). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.