Department of Bioengineering, Graduate School of Science and Technology, Hacettepe University, Ankara, Turkey.
Department of Molecular Biology and Genetics, Bilkent University, Ankara, Turkey.
PLoS One. 2023 Feb 28;18(2):e0282238. doi: 10.1371/journal.pone.0282238. eCollection 2023.
Mesenchymal stem cell-derived exosomes regulate cell migration, proliferation, differentiation, and synthesis of the extracellular matrix, giving great potential for the treatment of different diseases. The ultracentrifugation method is the gold standard method for exosome isolation due to the simple protocol, and high yield, but presents low purity and requires specialized equipment. Amelioration of technical optimization is required for quick and reliable confinement of exosomes to translate them to the clinic as cell therapeutics In this study, we hypothesized that magnetically activated cell sorting may provide, an effective, reliable, and rapid tool for exosome isolation when compared to ultracentrifugation. We, therefore, aimed to compare the efficiency of magnetically activated cell sorting and ultracentrifugation for human mesenchymal stem cell-derived exosome isolation from culture media by protein quantification, surface biomarker, size, number, and morphological analysis. Magnetically activated cell sorting provided a higher purity and amount of exosomes that carry visible magnetic beads when compared to ultracentrifugation. The particle number of the magnetically activated cell sorting group was higher than the ultracentrifugation. In conclusion, magnetically activated cell sorting presents a quick, and reliable method to collect and present human mesenchymal stem cell exosomes to clinics at high purity for potential cellular therapeutic approaches. The novel isolation and purification method may be extended to different clinical protocols using different autogenic or allogeneic cell sources.
间充质干细胞衍生的外泌体调节细胞迁移、增殖、分化和细胞外基质的合成，为治疗不同疾病提供了巨大的潜力。超速离心法是外泌体分离的金标准方法，因为其方案简单，产量高，但纯度低，需要专用设备。需要对技术进行优化改进，以便快速可靠地捕获外泌体，并将其作为细胞治疗方法转化为临床应用。在本研究中，我们假设与超速离心法相比，磁激活细胞分选可能为外泌体分离提供一种有效、可靠和快速的工具。因此，我们旨在通过蛋白质定量、表面标志物、大小、数量和形态分析比较磁激活细胞分选和超速离心法从培养介质中分离人骨髓间充质干细胞衍生的外泌体的效率。与超速离心法相比，磁激活细胞分选提供了更高纯度和数量的携带可见磁珠的外泌体。磁激活细胞分选组的颗粒数高于超速离心法。总之，磁激活细胞分选提供了一种快速、可靠的方法，以高纯度将人骨髓间充质干细胞外泌体收集并呈现给临床，用于潜在的细胞治疗方法。这种新的分离和纯化方法可以扩展到使用不同自体或同种异体细胞来源的不同临床方案。
