Pamulang Yudith Violetta, Oontawee Saranyou, Rodprasert Watchareewan, Padeta Irma, Sa-Ard-Lam Noppadol, Mahanonda Rangsini, Osathanon Thanaphum, Somparn Poorichaya, Pisitkun Trairak, Torsahakul Chutirat, Sawangmake Chenphop
The International Graduate Program of Veterinary Science and Technology (VST), Faculty of Veterinary Science, Chulalongkorn University, Bangkok, 10330, Thailand.
Center of Excellence for Veterinary Clinical Stem Cells and Bioengineering, Chulalongkorn University, Bangkok, 10330, Thailand.
Sci Rep. 2025 Mar 27;15(1):10617. doi: 10.1038/s41598-025-93219-7.
Mesenchymal stem cell-derived exosomes exhibit promising potential in tissue regeneration. Recent studies highlight its significant therapeutic potential in various stages of wound healing. However, the clinical translation of exosome-based therapy was hindered due to issues regarding low productivity and the lack of efficient production protocol to obtain a clinically relevant exosome quantity. Therefore, this study established a potential upscaling protocol to produce exosomes derived from canine adipose-derived mesenchymal stem cells (cAD-MSCs) and explored its potential for wound treatment. The potential upscaling protocol, termed VSCBIC-3-3D, was carried out using VSCBIC-3 in-house serum-free exosome-collecting solution in a three-dimensional (3D) culture system followed by the tangential flow filtration (TFF) isolation. Our findings suggest that culturing cAD-MSCs with VSCBIC-3 maintained cell morphology and viability. Compared to conventional two-dimensional (2D) protocols, The potential upscaling protocol increased exosome yield and concentration in conditioned medium by 2.4-fold and 3.2-fold, respectively. The quality assessment revealed enhanced purity and bioactivity of exosomes produced using the VSCBIC-3-3D protocol. In addition, the cAD-MSCs-derived exosomes were shown to significantly improve fibroblast migration, proliferation, and wound healing-related gene expression in vitro. This study collectively demonstrates that potential upscaling protocol establishment allowed robust production of exosomes from cAD-MSCs, which exhibit therapeutic potential for wound healing in vitro.
间充质干细胞衍生的外泌体在组织再生方面展现出了有前景的潜力。最近的研究突出了其在伤口愈合各个阶段的显著治疗潜力。然而，由于产量低以及缺乏获得临床相关外泌体数量的高效生产方案等问题，基于外泌体的疗法的临床转化受到了阻碍。因此，本研究建立了一种潜在的扩大规模方案来生产犬脂肪来源间充质干细胞（cAD-MSCs）衍生的外泌体，并探索其伤口治疗潜力。该潜在的扩大规模方案称为VSCBIC-3-3D，是在三维（3D）培养系统中使用VSCBIC-3内部无血清外泌体收集溶液进行，随后通过切向流过滤（TFF）分离。我们的研究结果表明，用VSCBIC-3培养cAD-MSCs可维持细胞形态和活力。与传统的二维（2D）方案相比，该潜在的扩大规模方案使条件培养基中外泌体的产量和浓度分别提高了2.4倍和3.2倍。质量评估显示，使用VSCBIC-3-3D方案生产的外泌体纯度和生物活性有所提高。此外，cAD-MSCs衍生的外泌体在体外显著改善了成纤维细胞的迁移、增殖以及与伤口愈合相关的基因表达。本研究共同表明，潜在扩大规模方案的建立使得能够从cAD-MSCs大量生产外泌体，这些外泌体在体外展现出伤口愈合的治疗潜力。
