Proteomics

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Altered extracellular matrix protein dynamics distinguishes canine ligament-derived stem cells from fibroblasts


ABSTRACT: Ligaments are prone to injury and degeneration in humans and animals, however the healing potential of ligament is poor and current treatment options lack effectiveness. Stem cell-based therapies hold potential for treatment of ligament injuries. This study aimed to characterise ligament-derived stem cell (LDSC) and ligament fibroblast populations as well as to identify specific niche components which could be used to promote the survival and function of LDSCs. LDSCs and ligament fibroblasts were isolated from canine cranial cruciate ligament and characterised for clonogenicity, multipotency and marker expression. The extracellular matrix (ECM) composition was characterised using a novel metabolic labelling and mass spectrometry approach. Both cell types demonstrated clonogenicity, with significant differences seen in colony size. Stem cell and tenogenic marker expression was increased in LDSCs compared with ligament fibroblasts, although this was only significant for the MSC marker CD73. Both cell types demonstrated multipotency, however this was more marked in LDSCs. A number of collagens, glycoproteins, proteoglycans and ECM regulators were identified in both the LDSC and ligament fibroblast niche using proteomics, with some differences between cell types, including collagens, decorin and fibronectin. Metabolic labelling of cells demonstrated unique turnover profiles for distinct ECM protein groups, with significant differences in new synthesis seen between cell types for a number of proteins, including collagen, decorin and elastin. In conclusion, different cell populations were observed in canine ligament with differences in niche composition and dynamics seen between the two cell types. These differences could be exploited to aid identification of LDSCs and to promote survival and function of LDSCs for potential therapeutic strategies.

INSTRUMENT(S): Q Exactive

ORGANISM(S): Canis Familiaris (dog) (canis Lupus Familiaris)

TISSUE(S): Primary Cell, Stem Cell, Fibroblast

DISEASE(S): Connective Tissue Disease

SUBMITTER: Katie Lee  

LAB HEAD: Dr Elizabeth Canty-Laird

PROVIDER: PXD008602 | Pride | 2019-01-28

REPOSITORIES: Pride

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Publications

Identification and Characterization of Canine Ligament Progenitor Cells and Their Extracellular Matrix Niche.

Lee Katie J KJ   Comerford Eithne J EJ   Simpson Deborah M DM   Clegg Peter D PD   Canty-Laird Elizabeth G EG  

Journal of proteome research 20190207 3


Ligaments are prone to injury and degeneration in humans and animals, however the healing potential of ligament is poor and current treatment options ineffective. Stem cell-based therapies hold potential for treatment of ligament injuries. This study aimed to characterize a ligament progenitor cell (LPC) population and to identify specific niche components which could promote the survival and function of LPCs. LPCs were isolated from canine cranial cruciate ligament and characterized for clonoge  ...[more]

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