Comparison of the Efficiency of Ultrafiltration, Precipitation, and Ultracentrifugation Methods for Exosome Isolation
Extracellular vesicles (EVs) are enclosed by a lipid-bilayer membrane and secreted by all types of the cells under various physiological conditions. EVs play important roles in intercellular communication and crosstalk between tissues in the body. They are classified into three groups, such as apoptotic bodies, microvesicles, and exosomes. Exosomes were isolated from biofluids including blood, urine, milk, and cell culture media. Exosomes have significant potential for drug delivery and diagnosis. However, the method of isolation affects the physical and biological properties of exosomes. Several methods based on different principles have been developed for exosome isolation. These include ultrafiltration, precipitation, ultracentrifugation, size-exclusion chromatography, and microfluidics. In this study, we applied three common methods, such as ultrafiltration, precipitation, and ultracentrifugation, to isolate exosomes from the cell culture medium and investigated the effects of these different isolation methods on the size distribution and quality of the isolated exosomes. Field emission scanning electron microscopy (FESEM) images, size distribution, total protein content, and the effect of exosomes on the viability of hypoxic cells were analyzed in this study. The analysis revealed that compared to other methods, the ultracentrifugation method can isolate exosomes with smaller diameter (ranging from 20 to 80 nm), lower total protein content (50 µg/ml), and causing the increased viability of the hypoxic cells. The precipitation method does not require special equipment and is inexpensive, if the quality and purity of this method are solved, and it can be used as the best method for exosome isolation. This study can serve as a guide for choosing the best exosome isolation method for applications in medicine according to the needs, time, cost, and equipment..
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Preprint| Erscheinungsjahr: |
2024 |
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| Erschienen: |
2024 |
| Enthalten in: |
Preprints.org - (2024) vom: 11. März Zur Gesamtaufnahme - year:2024 |
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| Sprache: |
Englisch |
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| Beteiligte Personen: |
Ansari, Farshid Jaberi [VerfasserIn] |
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| Links: |
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| doi: |
10.20944/preprints202308.1202.v2 |
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| PPN (Katalog-ID): |
preprintsorg040556417 |
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| 520 | |a Extracellular vesicles (EVs) are enclosed by a lipid-bilayer membrane and secreted by all types of the cells under various physiological conditions. EVs play important roles in intercellular communication and crosstalk between tissues in the body. They are classified into three groups, such as apoptotic bodies, microvesicles, and exosomes. Exosomes were isolated from biofluids including blood, urine, milk, and cell culture media. Exosomes have significant potential for drug delivery and diagnosis. However, the method of isolation affects the physical and biological properties of exosomes. Several methods based on different principles have been developed for exosome isolation. These include ultrafiltration, precipitation, ultracentrifugation, size-exclusion chromatography, and microfluidics. In this study, we applied three common methods, such as ultrafiltration, precipitation, and ultracentrifugation, to isolate exosomes from the cell culture medium and investigated the effects of these different isolation methods on the size distribution and quality of the isolated exosomes. Field emission scanning electron microscopy (FESEM) images, size distribution, total protein content, and the effect of exosomes on the viability of hypoxic cells were analyzed in this study. The analysis revealed that compared to other methods, the ultracentrifugation method can isolate exosomes with smaller diameter (ranging from 20 to 80 nm), lower total protein content (50 µg/ml), and causing the increased viability of the hypoxic cells. The precipitation method does not require special equipment and is inexpensive, if the quality and purity of this method are solved, and it can be used as the best method for exosome isolation. This study can serve as a guide for choosing the best exosome isolation method for applications in medicine according to the needs, time, cost, and equipment. | ||
| 700 | 1 | |a Tafti, Hossein Ahmadi |4 aut | |
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