Department of Orthopedics, The First affiliated hospital of Shenzhen University, Shenzhen Second People's Hospital; Guangdong Provincial Research Center for Artificial Intelligence and Digital Orthopedic Technology, Shenzhen Second People's Hospital.
Department of Orthopedics, The First affiliated hospital of Shenzhen University, Shenzhen Second People's Hospital; Guangdong Provincial Research Center for Artificial Intelligence and Digital Orthopedic Technology, Shenzhen Second People's Hospital;
J Vis Exp. 2022 Jul 26(185). doi: 10.3791/62221.
Exosomes secreted by mesenchymal stem cells (MSCs) have been suggested as promising candidates for cartilage injuries and osteoarthritis treatment. Exosomes for clinical application require large-scale production. To this aim, human synovial fluid MSCs (hSF-MSCs) were grown on microcarrier beads, and then cultured in a dynamic three-dimension (3D) culture system. Through utilizing 3D dynamic culture, this protocol successfully obtained large-scale exosomes from SF-MSC culture supernatants. Exosomes were harvested by ultracentrifugation and verified by a transmission electron microscope, nanoparticle transmission assay, and western blotting. Also, the microbiological safety of exosomes was detected. Results of exosome detection suggest that this approach can produce a large number of good manufacturing practices (GMP)-grade exosomes. These exosomes could be utilized in exosome biology research and clinical osteoarthritis treatment.
间充质干细胞（MSCs）分泌的细胞外囊泡已被认为是治疗软骨损伤和骨关节炎的有希望的候选物。用于临床应用的细胞外囊泡需要大规模生产。为此，在微载体珠上培养人滑膜间充质干细胞（hSF-MSCs），然后在动态三维（3D）培养系统中培养。通过利用 3D 动态培养，该方案成功地从 SF-MSC 培养上清液中获得了大量的细胞外囊泡。通过超速离心法收获细胞外囊泡，并通过透射电子显微镜、纳米颗粒传输分析和 Western blot 进行验证。此外，还检测了细胞外囊泡的微生物安全性。细胞外囊泡检测结果表明，该方法可以产生大量良好生产规范（GMP）级别的细胞外囊泡。这些细胞外囊泡可用于细胞外囊泡生物学研究和临床骨关节炎治疗。
