Abstract
BackgroundThe dog represents an excellent large animal model for translational cell-based studies. Importantly, the properties of canine multipotent stromal cells (cMSCs) and the ideal tissue source for specific translational studies have yet to be established. The aim of this study was to characterize cMSCs derived from synovium, bone marrow, and adipose tissue using a donor-matched study design and a comprehensive series of in-vitro characterization, differentiation, and immunomodulation assays.MethodsCanine MSCs were isolated from five dogs with cranial cruciate ligament rupture. All 15 cMSC preparations were evaluated using colony forming unit (CFU) assays, flow cytometry analysis, RT-PCR for pluripotency-associated genes, proliferation assays, trilineage differentiation assays, and immunomodulation assays. Data were reported as mean ± standard deviation and compared using repeated-measures analysis of variance and Tukey post-hoc test. Significance was established at p < 0.05.ResultsAll tissue samples produced plastic adherent, spindle-shaped preparations of cMSCs. Cells were negative for CD34, CD45, and STRO-1 and positive for CD9, CD44, and CD90, whereas the degree to which cells were positive for CD105 was variable depending on tissue of origin. Cells were positive for the pluripotency-associated genes NANOG, OCT4, and SOX2. Accounting for donor and tissue sources, there were significant differences in CFU potential, rate of proliferation, trilineage differentiation, and immunomodulatory response. Synovium and marrow cMSCs exhibited superior early osteogenic activity, but when assessing late-stage osteogenesis no significant differences were detected. Interestingly, bone morphogenic protein-2 (BMP-2) supplementation was necessary for early-stage and late-stage osteogenic differentiation, a finding consistent with other canine studies. Additionally, synovium and adipose cMSCs proliferated more rapidly, displayed higher CFU potential, and formed larger aggregates in chondrogenic assays, although proteoglycan and collagen type II staining were subjectively decreased in adipose pellets as compared to synovial and marrow pellets. Lastly, cMSCs derived from all three tissue sources modulated murine macrophage TNF-α and IL-6 levels in a lipopolysaccharide-stimulated coculture assay.ConclusionsWhile cMSCs from synovium, marrow, and adipose tissue share a number of similarities, important differences in proliferation and trilineage differentiation exist and should be considered when selecting cMSCs for translational studies. These results and associated methods will prove useful for future translational studies involving the canine model.
Highlights
The dog represents an excellent large animal model for translational cell-based studies
While Canine Multipotent stromal cell (MSC) (cMSC) from synovium, marrow, and adipose tissue share a number of similarities, important differences in proliferation and trilineage differentiation exist and should be considered when selecting cMSCs for translational studies
Tumor necrosis factor alpha (TNF-α) concentrations were significantly decreased in cocultures containing 5 × 104 cells (p < 0.0001) and 2.5 × 104 cells (p < 0.01) when compared to cocultures containing 1 × 103 cells/well. These results suggest that cMSCs from synovium, marrow, and adipose tissues are capable of Interestingly, while TNF-α concentrations decreased in response to cMSC coculture, the concentration of murine Interleukin 6 (IL-6) increased in the LPS coculture assay (Additional file 3)
Summary
The dog represents an excellent large animal model for translational cell-based studies. Translation of promising findings from rodent models to humans represents a significant hurdle for cell-based therapies For this reason, a number of large animal species have been used to bridge the gap from rodents to humans [1,2,3,4]. Humans often consider dogs as in-home pets, exposing both to similar environmental stimuli, which helps to eliminate variables between species [20, 21] For many of these reasons, canine spontaneous diseases have been used to translate novel therapeutics to humans [22, 23]. Benefits of the canine translational model have been described extensively in recent review articles [24, 25]
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a call
