Comparison of equine tendon-, muscle-, and bone marrow–derived cells cultured on tendon matrix

Abstract

Objective-To compare viability and biosynthetic capacities of cells isolated from equine tendon, muscle, and bone marrow grown on autogenous tendon matrix. Sample Population-Cells from 4 young adult horses. Procedures-Cells were isolated, expanded, and cultured on autogenous cell-free tendon matrix for 7 days. Samples were analyzed for cell viability, proteoglycan synthesis, collagen synthesis, and mRNA expression of collagen type I, collagen type III, and cartilage oligomeric matrix protein (COMP). Results-Tendon-and muscle-derived cells required less time to reach confluence (approx 2 weeks) than did bone marrow-derived cells (approx 3 to 4 weeks); there were fewer bone marrow-derived cells at confluence than the other 2 cell types. More tendon-and musclederived cells were attached to matrices after 7 days than were bone marrow-derived cells. Collagen and proteoglycan synthesis by tendon-and muscle-derived cells was significantly greater than synthesis by bone marrow-derived cells. On a per-cell basis, tendon-derived cells had more collagen synthesis, although this was not significant. Collagen type I mRNA expression was similar among groups. Tendon-derived cells expressed the highest amounts of collagen type III and COMP mRNAs, although the difference for COMP was not significant. Conclusions and Clinical Relevance-Tendon-and muscle-derived cells yielded greater cell culture numbers in shorter time and, on a per-cell basis, had comparable biosynthetic assays to bone marrow-derived cells. More in vitro experiments with higher numbers may determine whether tendon-derived cells are a useful resource for tendon healing. (Am J Vet Res 2009;70:750-757)

Freeze-Thaw Cycles Enhance Decellularization of Large Tendons

“…25 However, our own unpublished data showed that this procedure, although reducing the content of vital cells, did not remove DNA, which was recently supported by others. 18 Freeze-thaw cycles with subsequent nuclease treatment led to nearly complete removal of cells and DNA, 18 but nuclease treatment was performed on sliced tendon only, 18,21 thus, this protocol does not appear applicable for decellularization of whole, large tendon constructs.…”

Cell- and gene-based approaches to tendon regeneration

“…29,59,75 Other autologous cell sources, including adipose tissue, muscle, synovium, periosteum, tendon-derived stem cells, dermal fibroblasts, and stem cells derived from umbilical cord or peripheral blood, have all been evaluated as sources of multipotent and pluripotent cells. 19,97,120,129,141,144 Adipose-derived stem cells are relatively easy to harvest, and selection of adherent cells over the course of several weeks in monolayer culture provides a cell type that improves the healing potential of flexor tendons. 129 Similarly, stem cells derived from skeletal muscle have been used in a variety of musculoskeletal repair indications and are readily harvested and propagated before reimplantation.…”

Aligned Nanofiber Topography Directs the Tenogenic Differentiation of Mesenchymal Stem Cells

“…MSCs were isolated from bone marrow aspirate harvested from the sternum of a one year old male horse, euthanized for unrelated conditions, following approval of the Institutional Animal Care and Use Committee (IACUC) of the Virginia Polytechnic Institute and State University and assessed via previously described techniques [28]. Briefly, bone marrow was collected, centrifuged to remove the plasma, and cultured in media containing low-glucose GlutaMAX DMEM with 110 mg/mL sodium pyruvate (Thermo Fisher Scientific, Waltham, MA, USA), supplemented with 10% MSCs fetal bovine serum (Thermo Fisher Scientific, Waltham, MA, USA), and 1% sodium penicillin and streptomycin sulfate (Sigma, St. Louis, MO, USA) at 37 • C, 5% CO 2 and 90% humidity.…”