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Comparison of two extraction methods of extracellular vesicles from Clostridium perfringens
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Soheila Ostadmohammadi1    , Seyed Ali Nojoumi 2, Abolfazl Fateh3 , Seyed Davar Siadat4 , Fattah Sotoodehnejadnematalahi5 |
1- PhD Student in Microbiology, Department of Biology, Science and Research Branch, Islamic Azad University, Tehran, Iran
2- Assistant Professor of Microbiology, Department of Mycobacteriology and Pulmonary Research, Pasteur Institute of Iran, Tehran, Iran , ali.nojoumi@gmail.com
3- Associate Professor of Medical Virology, Department of Mycobacteriology and Pulmonary Research, Pasteur Institute of Iran, Tehran, Iran
4- Professor of Medical Bacteriology, Department of Mycobacteriology and Pulmonary Research, Pasteur Institute of Iran, Tehran, Iran
5- Assistant Professor of Cellular and Molecular Immunology, Department of Biology, Science and Research Branch, Islamic Azad University, Tehran, Iran |
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Abstract: (615 Views) |
Background: Extracellular vesicles (EVs) are nano- to micron-sized vesicles with the ability to transport bioactive cargos. All types of cells have the ability to release EVs, which have been demonstrated to be involved in a number of essential cell functions. There are various methods for isolating EVs, each with advantages and disadvantages. The purpose of this study was to invistigate at the non-ultra centrifugation method and ultra-centrifugation method for isolating EVs.
Materials and methods: Clostridium perfringens ATCC13124 was used in this experimental study. Following culture, EVs were extracted using two methods: Non-Ultra method and Ultra-method. To examine chemical characteristics, the EV protein concentration was measured using a NanoDrop device, and the EV protein pattern was identified using SDS-PAGE. Transmission electron microscopy (TEM) was utilized to examine the EVs' physical properties.
Results: Our results showed that the EVs isolated by the Ultra-method had a higher protein content compared to the Non-Ultra method (3.17 and 1.46 mg/ml, respectively).The Ultra-method isolated more and larger EVs compared to the Non-Ultra method. Also, protein patterns of the EVs by SDS-PAGE method were similar in both methods.
Conclusion: The present study showed that the Ultra-method can be a more efficient and cost-effective way for isolation EVs from Clostridium perfringens than the Non-Ultra method.
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| Keywords: Clostridium perfringens, Extracellular vesicles, ultracentrifugation, EV extraction methods |
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Full-Text [PDF 800 kb]
(216 Downloads)
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Semi-pilot: Experimental |
Subject:
Microbiology
Received: 2023/08/19 | Accepted: 2023/09/26 | Published: 2024/06/30
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Ostadmohammadi S, Nojoumi S A, Fateh A, Siadat S D, Sotoodehnejadnematalahi F. Comparison of two extraction methods of extracellular vesicles from Clostridium perfringens. MEDICAL SCIENCES 2024; 34 (2) :121-128 URL: http://tmuj.iautmu.ac.ir/article-1-2160-en.html
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