Abstract
ABSTRACTThe aim of this study was to characterize stem and progenitor cell populations from the equine superficial digital flexor tendon, an energy‐storing tendon with similarities to the human Achilles tendon, which is frequently injured. Using published methods for the isolation of tendon‐derived stem/progenitor cells by low‐density plating we found that isolated cells possessed clonogenicity but were unable to fully differentiate towards mesenchymal lineages using trilineage differentiation assays. In particular, adipogenic differentiation appeared to be restricted, as assessed by Oil Red O staining of stem/progenitor cells cultured in adipogenic medium. We then assessed whether differential adhesion to fibronectin substrates could be used to isolate a population of cells with broader differentiation potential. However we found little difference in the stem and tenogenic gene expression profile of these cells as compared to tenocytes, although the expression of thrombospondin‐4 was significantly reduced in hypoxic conditions. Tendon‐derived stem/progenitor cells isolated by differential adhesion to fibronectin had a similar differentiation potential to cells isolated by low density plating, and when grown in either normoxic or hypoxic conditions. In summary, we have found a restricted differentiation potential of cells isolated from the equine superficial digital flexor tendon despite evidence for stem/progenitor‐like characteristics. © 2015 The Authors. Journal of Orthopaedic Research Published by Wiley Periodicals, Inc. on behalf of Orthopaedic Research Society. J Orthop Res 33:849–858, 2015.
Highlights
Sports participation, occupation, and aging increase the risk of tendon injury and degeneration in both humans and animals.[1,2,3] In the horse, the superficial digital flexor tendon (SDFT) is commonly injured, resulting in lameness and reduced performance, in athletic and racing horses.[3]
Tendon-derived stem/ progenitor cells (TSPCs) from Equine SDF Tendon form Colonies when Plated at Low Density but have Restricted Trilineage differentiation TSPCs isolated from SDF tendon by low-density plating were able to form colonies after replating, as were tenocytes initially plated at high density (Fig. 1A–C)
For TSPCs grown in osteogenic media, there were no significant differences in the expression of osteogenic markers runx[2] (RUNX2), alkaline phosphatase (ALP), osteocalcin (BGLAP), or osteomodulin (OMD) in osteogenic as compared to control media (Fig. 2M)
Summary
Occupation, and aging increase the risk of tendon injury and degeneration in both humans and animals.[1,2,3] In the horse, the superficial digital flexor tendon (SDFT) is commonly injured, resulting in lameness and reduced performance, in athletic and racing horses.[3]. The identification of a population of cells within tendon with stem cell-like characteristics[8] holds poten-. Tendon-derived stem/ progenitor cells (TSPCs) have been identified in human, mouse,[8] rat,[9] and rabbit tendon.[10] TSPCs possess similar properties to mesenchymal stem cells (MSCs) and have been identified by the expression of cell surface and stem-cell markers, and a capacity for selfrenewal and multi-lineage differentiation. TSPCs are thought to be tenocyte precursors and can be induced to differentiate into osteocytes, chondrocytes, and adipocytes in vitro and in vivo.[8,9,10,11,12]
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