Abstract
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 clonogenicity, multipotency and marker expression. The extracellular matrix (ECM) composition was characterized by the novel application of a metabolic labeling and mass spectrometry technique. LPCs demonstrated clonogenicity, multipotency, and stem cell marker expression. A number of different collagens, glycoproteins, and proteoglycans were identified in the LPC niche using proteomics. Metabolic labeling of cells demonstrated unique turnover profiles for distinct ECM protein groups, indicating the importance of certain niche components for LPC survival and function. The newly synthesized niche components identified in this study could be exploited to aid identification of LPCs and to promote their survival and function for potential ligament repair strategies.
Highlights
Musculoskeletal soft tissues such as ligament are primarily composed of extracellular matrix (ECM) within which ligament cell populations reside
ligament progenitor cell (LPC) Display Clonogenicity and Stem Cell Marker Expression Ligament-derived progenitor cells grew in heterogeneous dense colonies (Figure 1A−C.) and showed a rounded and fibroblastic morphology upon initial plating (Figure 1D.), with the rounded morphology lost with passaging
Our findings are consistent with previous studies in humans, with stem/progenitor cells isolated from human anterior cruciate ligament (ACL) demonstrating the same properties as the cells isolated in this study.[18−20] Despite the stem cell properties demonstrated by the cell population selectively isolated in this study, it is likely that this is a heterogeneous population of cells, as clonal cell populations were not derived and different cell morphologies were observed upon initial plating
Summary
Musculoskeletal soft tissues such as ligament are primarily composed of extracellular matrix (ECM) within which ligament cell populations reside. These tissues are prone to injury and degeneration, the anterior cruciate ligament (ACL),[1] with an incidence of approximately 37 ACL ruptures per 100 000 people[2] and a greater incidence among athletes.[3]. Rupture of the cranial cruciate ligament (CCL), comparable to the human ACL, is the predominant cause of canine hind limb lameness.[8] Study of the canine CCL is important for its translation into humans as a model for ACL disease.[9,10] Current treatment strategies for human ACL and canine CCL injuries have variable success rates,[11−16] a more effective therapeutic option for treatment of ACL and CCL injury is currently being sought
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