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概要

Platelet-Rich Plasma Enhances the Cellular Function of Equine Bone Marrow-Derived Mesenchymal Stem Cells

Madhu Dhar, Lisa Amelse, Nancy Neilsen, Pelagie Favi and Jessica Carter-Arnold

Rationale: Equine mesenchymal stem cells (eMSCs) and platelet-rich plasma (PRP) are cell-based therapies being used clinically to repair damaged tissue of horses. Numerous reports including data from our laboratory show that there are variations in the biological properties of MSCs and platelet – rich plasma, which can impact their biological functions. A single study describes the use of the eMSCs and PRP in tendon healing, with minimal success. The exact mechanism of action using the combination of the two therapies is unknown. This study was performed to evaluate and understand the effects of PRP, if any on the cellular performance of eBMMSCs in culture.
Objective: To assess the effects of PRP in vitro on the rate of proliferation, expression of protein markers and osteogenic and chondrogenic differentiation of the primary cultures of eBMMSCs.
Methods and Results: A commercially available stall side, portable kit was used to isolate PRP. To investigate the effect of PRP on eBMMSCs, the rate of proliferation of eBMMSCs was measured using the MTS assay, and subsequently the viability and the stemness of eBMMSCs was assessed using fluorescent staining and the expression of CD90. Finally, the osteogenic and chondrogenic differentiation of eBMMSCs was assessed by lineage-specific staining and the expressions of lineage – specific mRNAs. All assays were performed at a concentration of 50 million platelets/mL. Significant increase in proliferation and the differentiation profiles were observed in presence of PRP. Most importantly, the stem cell characteristics of inferior eBMMSCs showed a marked improvement in presence of PRP.
Conclusions: The addition of PRP improves the in vitro function of eBMMSCs by enhancing the proliferation and osteo - and chondro - genesis. The presence of an optimal dose of platelets may enhance the in vivo performance of eBMMSCs and may be indicated when using autologous eBMMSCs therapy in the clinic.

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