Characterization of the secretory profile and exosomes of limbal stem cells in the canine species

PLoS One. 2020 Dec 29;15(12):e0244327. doi: 10.1371/journal.pone.0244327. eCollection 2020.

Abstract

Limbal stem cells (LSCs) are a quiescent cell population responsible for the renewal of the corneal epithelium. Their deficiency is responsible for the conjunctivization of the cornea that is seen in different ocular pathologies, both in humans and in the canine species. The canine species represents an interesting preclinical animal model in ocular surface pathologies. However, the role of LSCs in physiological and pathological conditions in canine species is not well understood. Our objective was to characterize for the first time the soluble factors and the proteomic profile of the secretome and exosomes of canine LSCs (cLSCs). In addition, given the important role that fibroblasts play in the repair of the ocular surface, we evaluated the influence of the secretome and exosomes of cLSCs on their proliferation in vitro. Our results demonstrated a secretory profile of cLSCs with high concentrations of MCP-1, IL-8, VEGF-A, and IL-10, as well as significant production of exosomes. Regarding the proteomic profile, 646 total proteins in the secretome and 356 in exosomes were involved in different biological processes. Functionally, the cLSC secretome showed an inhibitory effect on the proliferation of fibroblasts in vitro, which the exosomes did not. These results open the door to new studies on the possible use of the cLSC secretome or some of its components to treat certain pathologies of the ocular surface in canine species.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Biomarkers / metabolism*
  • Cell Culture Techniques
  • Cell Differentiation
  • Cell Proliferation
  • Cells, Cultured
  • Chemokine CCL2 / metabolism
  • Culture Media / analysis*
  • Dogs
  • Exosomes / metabolism*
  • Interleukin-10 / metabolism
  • Interleukin-8 / metabolism
  • Limbus Corneae / cytology*
  • Limbus Corneae / metabolism
  • Proteomics / methods*
  • Stem Cells / cytology
  • Stem Cells / metabolism
  • Vascular Endothelial Growth Factor A / metabolism

Substances

  • Biomarkers
  • Chemokine CCL2
  • Culture Media
  • Interleukin-8
  • Vascular Endothelial Growth Factor A
  • Interleukin-10

Grants and funding

GR was supported by the RD16/0011/0022 grant of the Spanish Network on Cell Therapy (TerCel). AV and VF were supported by the Contract Aljama Gestión SL-University of Malaga (8.06/5.02.5013). MM was supported by the UMA18-FEDERJA-133 grant of the Operational programme FEDER Andalucía 2014-2020. CA was supported by the Promotion of Youth Employment and implementation of the Youth Guarantee in R&D&I, 2018 (PEJ2018-004785-A) grant of the Ministry of Science, Innovation and University. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. VI National R&D&I Plan 2008–2011, Iniciativa Ingenio 2010, Consolider Program, CIBER Actions, Instituto de Salud Carlos III with assistance from the European Regional Development Fund and Regional Government of Andalusia (Junta de Andalucia, PAIDI (BIO217), provided support in the form of materials, but did not have any additional role in the study design, data Cover Letter collection and analysis, decision to publish, or preparation of the manuscript. There was no additional external funding received for this study.