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Exosomes are a form of extracellular vesicle (EV), which could contribute to malignant transformation and the metastasis of cancer. Consequently, intercellular communication via exosomes is attracting considerable interest in the scientific community.
The purity of isolated exosomes and the amount of contamination are important factors that significantly impact the results of subsequent experiments.
The Isolation Filter is also sold separately in a pack of 10 pieces :
Ultracentrifugation is the most commonly used method to isolate exosomes. We isolated the exosomes from the supernatant of HEK293S using both of ultracentrifugation method and the ExoIsolator method. The particle size distribution (Fig. 1), the number of particles (Fig. 2(a)) and the expression level of exosome markers (Fig. 2(b)) of the isolated exosomes were tested and compared.
Exosomes stained with ExoSparkler’s Mem Dye-Deep Red or an alternative product (green or red) were added to each well containing HeLa cells. The labeled exosomes taken into HeLa cells were observed by fluorescent microscopy. As a result, extracellular fluorescent spots suspected of dye aggregations were seen in each well containing the exosomes stained with the alternative product (green or red).
NTA (nanoparticle tracking analysis) is measured to determine the changes in exosomes of dye-stained with Mem Dye-Deep Red or Product “P” (green or red) or unstained exosomes. As a result, the Mem-Dye series (green, red, deep red) had little effect on exosome properties.
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H. Chuang, M. Chen, Y. Chen, Y. Ciou, C. Hsueh, C. Tsai and T. Tan, “Induction of Interferon-γ and Tissue Inflammation by Overexpression of Eosinophil Cationic Protein in T Cells and Exosomes”, 2021, doi:10.1002/art.41920.
N. Kamei, H. Nishimura, A. Matsumoto, R. Asano, K. Muranaka, M. Fujita, M. Takeda, H. Hashimoto, M. Takeda-Morishita, “Comparative study of commercial protocols for high recovery of high-purity mesenchymal stem cell-derived extracellular vesicle isolation and their efficient labeling with fluorescent dyes,”, 2021, doi:10.1016/j.nano.2021.102396.
H. Chuang, M. Chen, Y. Chen, H. Yang, Y. Ciou, C. Hsueh, C. Tsai, T. Tan, “BPI overexpression suppresses Treg differentiation and induces exosome-mediated inflammation in systemic lupus erythematosus”, 2021, doi:10.7150/thno.63743.
H. Takakura, T. Nakao, T. Narita, M. Horinaka, Y. Nakao-Ise, T. Yamamoto, Y. Iizumi, M. Watanabe, Y. Sowa, K. Oda, N. Mori, T. Sakai, and M. Mutho, “Citrus limon L.-Derived Nanovesicles Show an Inhibitory Effect on Cell Growth in p53-Inactivated Colorectal Cancer Cells via the Macropinocytosis Pathway”, 2022, doi:10.3390/biomedicines10061352.
A. Shimoda R. Miura, H. Tateno, N. Seo, H. Shiku, S. Sawada, Y. Sasaki and K. Akiyoshi, “Assessment of Surface Glycan Diversity on Extracellular Vesicles by Lectin Microarray and Glycoengineering Strategies for Drug Delivery Applications”, Small Methods, 2021, doi:10.1002/smtd.202100785.
H. Takakura, T. Nakao, T. Narita, M. Horinaka, Y. Nakao-Ise, T. Yamamoto, Y. Iizumi, M. Watanabe, Y. Sowa, K. Oda, N. Mori, T. Sakai, and M. Mutho, “Citrus limon L.-Derived Nanovesicles Show an Inhibitory Effect on Cell Growth in p53-Inactivated Colorectal Cancer Cells via the Macropinocytosis Pathway”, 2022, doi:10.3390/biomedicines10061352.
Y. Wang, A. Chen, “Mast cell-derived exosomal miR-181a-5p modulated trophoblast cell viability, migration, and invasion via YY1/MMP-9 axis”, J. Clin. Lab. Anal., 2022, doi:10.1002/jcla.24549.
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