Vachkova Ekaterina, Arnhold Stefan, Petrova Valeria, Heimann Manuela, Koynarski Tsvetoslav, Simeonova Galina, Piperkov Paskal
Department of Pharmacology, Animal Physiology, Biochemistry and Chemistry, Faculty of Veterinary Medicine, Trakia University, 6000 Stara Zagora, Bulgaria.
Institute of Veterinary Anatomy, Histology and Embryology, Justus-Liebig-University of Giessen, 35392 Giessen, Germany.
Animals (Basel). 2025 Jun 28;15(13):1910. doi: 10.3390/ani15131910.
The impact of donor age on Adipose-derived mesenchymal stem cell (ASC) functionality and safety remains insufficiently characterized, particularly in equine models. This study investigates the influence of age on ASCs proliferation dynamics and the expression of tumorigenic and apoptosis-related markers. Equine ASCs were isolated from juvenile (<5 years), middle-aged (5-15 years), and geriatric (>15 years) horses and assayed across multiple passages. The relative mRNA expressions of pluripotency (Oct4), tumorigenic (CA9), and apoptosis-related (Bax and Bcl 2) markers were evaluated. The Gompertz growth model, population doubling time (PDT), and tissue non-specific ALP activity also followed. The expression of pluripotency and tumorigenic markers showed passage-dependent up-regulation, raising concerns about prolonged culture expansion. Apoptotic regulation displayed a shift with aging, as evidenced by alterations in the Bax/Bcl2 ratio, suggesting compromised cell survival in older ASCs. An age-associated decline in proliferation rates was established, as evidenced by declining alkaline phosphatase (ALP) activity. These findings underscore the necessity for stringent age-based selection criteria in equine stem cell therapies and the challenges associated with using autologous stem cells for regenerative therapies in aged horses. Future research should focus on molecular interventions to mitigate age-related functional decline, ensuring the safety and efficacy of ASCs-based regenerative medicine in equine practice.
供体年龄对脂肪间充质干细胞（ASC）功能和安全性的影响尚未得到充分研究，尤其是在马模型中。本研究调查了年龄对ASC增殖动力学以及致瘤和凋亡相关标志物表达的影响。从幼年（<5岁）、中年（5 - 15岁）和老年（>15岁）马中分离出ASC，并在多个传代中进行检测。评估了多能性（Oct4）、致瘤性（CA9）和凋亡相关（Bax和Bcl 2）标志物的相对mRNA表达。还采用了Gompertz生长模型、群体倍增时间（PDT）和组织非特异性碱性磷酸酶（ALP）活性进行分析。多能性和致瘤性标志物的表达显示出传代依赖性上调，这引发了对长期培养扩增的担忧。凋亡调节随着衰老而发生变化，Bax/Bcl2比值的改变证明了这一点，表明老年ASC的细胞存活能力受损。碱性磷酸酶（ALP）活性下降证明了增殖率随年龄下降。这些发现强调了在马干细胞治疗中基于年龄的严格选择标准的必要性，以及在老年马中使用自体干细胞进行再生治疗所面临的挑战。未来的研究应集中在分子干预措施上，以减轻与年龄相关的功能衰退，确保基于ASC的再生医学在马医学实践中的安全性和有效性。
