Poster Filip Humenik fin.pdf

Adult stem cells have become prominent candidates for treating various diseases in veterinary practice. The main goal of our study was therefore to provide a comprehensive study of canine bone marrow-derived mesenchymal stem cells (BMMSC) and conditioned media, isolated from healthy adult dogs of different breeds. Under well-defined standardized isolation protocols the multipotent differentiation and specific surface markers of MSCs were supplemented with their gene expression, proteomic profile and their biological function. The presented data confirm that canine BMMSC express important genes for differentiation toward osteo-, chondro- and tendo-genic directions, but also genes associated with angiogenic, neurotrophic and immunomodulatory properties. Furthermore, using proteome profiling we identify for the first time the dynamic release of various bioactive molecules, such as transcription and translation factors, osteogenic, growth, angiogenic and neurotrophic factors from canine BMMSC conditioned medium. Importantly, the relevant genes were linked to their proteins as detected in the conditioned medium, and further associated with angiogenic activity in chorioallantoic membrane (CAM) assay. In this way we show that the canine BMMSC release a variety of bioactive molecules revealing a strong paracrine component which may possess therapeutic potential in various pathologies. However, extensive experimental or pre-clinical trials testing canine sources need to be performed in order to better understand their paracrine action, which may lead to novel therapeutic strategies in veterinary medicine.

成体干细胞（Adult stem cells）现已成为兽医临床中治疗多种疾病的热门候选疗法。本研究的核心目标，是对从不同品种健康成年犬体内分离得到的犬源骨髓间充质干细胞（BMMSC）及其条件培养基开展全面系统的研究。我们采用规范统一的分离培养流程，在检测间充质干细胞（MSCs）多向分化潜能与特异性表面标志物的基础上，进一步补充了其基因表达特征、蛋白质组谱及生物学功能的相关研究。本研究所得数据证实，犬源BMMSC不仅表达向成骨、成软骨及成腱方向分化的关键基因，同时也携带与血管生成、神经营养及免疫调节特性相关的功能基因。此外，我们首次通过蛋白质组分析，鉴定出犬源BMMSC条件培养基中动态释放的多种生物活性分子，包括转录翻译因子、成骨相关因子、生长因子、血管生成因子及神经营养因子。值得注意的是，本研究将条件培养基中检测到的相关蛋白与其编码基因进行了关联，并进一步通过鸡胚绒毛尿囊膜（chorioallantoic membrane, CAM）实验证实其具备血管生成活性。综上，本研究证实犬源BMMSC可释放多种生物活性分子，展现出极强的旁分泌特性，这为其在多种病理状态下的治疗应用提供了潜力。但未来仍需开展针对犬源干细胞的大规模实验及临床前研究，以深入阐明其旁分泌作用机制，进而为兽医学领域开发全新的治疗策略奠定基础。