Evaluation of canine adipose-derived multipotent stromal cell differentiation to ligamentoblasts on tensioned collagen type I templates in a custom bioreactor culture system
OBJECTIVE: To evaluate differentiation of canine adipose-derived multipotent stromal cells (ASCs) into ligamentoblasts on tensioned collagen type I (Col1) templates in a perfusion culture system. SAMPLES: Infrapatellar fat pad ASCs from healthy stifle joints of 6 female mixed-breed dogs. PROCEDURES: Third-passage ASCs (6 × 10 cells/template) were loaded onto suture-augmented Col1 templates under 15% static strain in perfusion bioreactors. Forty-eight ASC-Col1 constructs were incubated with ligamentogenic (ligamentogenic constructs; n = 24) or stromal medium (stromal constructs; 24) for up to 21 days. Specimens were collected from each construct after 2 hours (day 0) and 7, 14, and 21 days of culture. Cell number, viability, distribution, and morphology; construct collagen content; culture medium procollagen-I-N-terminal peptide concentration; and gene expression were compared between ligamentogenic and stromal constructs. RESULTS: ASCs adhered to collagen fibers. Cell numbers increased from days 0 to 7 and days 14 to 21 for both construct types. Relative to stromal constructs, cell morphology and extracellular matrix were more mature and collagen content on day 21 and procollagen-I-N-terminal peptide concentration on days 7 and 21 were greater for ligamentogenic constructs. Ligamentogenic constructs had increased expression of the genes biglycan on day 7, decorin throughout the culture period, and , tenomodulin, fibronectin, and tenascin-c on day 21; expression of , tenomodulin, and tenascin-c increased between days 7 and 21. CONCLUSIONS AND CLINICAL RELEVANCE: Ligamentogenic medium was superior to stromal medium for differentiation of ASCs to ligamentoblasts on suture-augmented Col1 scaffolds. Customized ligament neotissue may augment treatment options for dogs with cranial cruciate ligament rupture.
Publication Source (Journal or Book title)
American journal of veterinary research
Taguchi, T., Zhang, N., Angibeau, D., Spivey, K. P., & Lopez, M. J. (2021). Evaluation of canine adipose-derived multipotent stromal cell differentiation to ligamentoblasts on tensioned collagen type I templates in a custom bioreactor culture system. American journal of veterinary research, 82 (11), 924-934. https://doi.org/10.2460/ajvr.82.11.924