Isolation and Characterization of Adipose-Derived Mesenchymal Stem Cell Exosomes: an In-Vitro Study

J. Fiolin, I. H. Dilogo, R. Antarianto, Ludwig Andre Pontoh
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引用次数: 1

Abstract

Exosomes from Mesenchymal Stem Cells (MSC) is currently one of the highlighted research due to its more specific action on the target tissue. Exosome is one of the MSC Extracellular Vesicle (EVs) that acts as a mediator in the cellular communication. However, not so many literatures have succeed in elaborating its isolation and characterization process. This study aimed to explain the method used in exosomes isolation from Adipose-Derived MSC (ASC) and elaborate its characteristics. This is an in-vitro study performed in the Stem Cell and Tissue Engineering (SCTE) Indonesia Medical Education and Research Institute (IMERI) laboratory. Ultracentrifugation method was performed to isolate exosome from ASC. The exosome were characterized based on its particle size, morphology, and CD63 and CD81 expression for its purity. We were able to isolate sterile exosome from ASC by performing differential ultracentrifugation. The mean size of exosome was 88.7 nm ± 40 nm SD and showed expression of CD63 and CD81. Exosome was successfully isolated from ASC using ultracentrifugation method and characterization following the MISEV standard should be implemented in order to meet the therapeutic efficacy and safety issues as a regenerative agent.
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脂肪来源的间充质干细胞外泌体的分离和表征:一项体外研究
来自间充质干细胞(MSC)的外泌体是目前重点研究的对象之一,因为它对靶组织具有更特异的作用。外泌体是MSC细胞外囊泡(EVs)中的一种,在细胞通信中起中介作用。然而,并没有那么多的文献成功地阐述了它的分离和表征过程。本研究旨在解释从脂肪衍生MSC(ASC)中分离外泌体的方法,并阐述其特征。这是一项在干细胞和组织工程(SCTE)印度尼西亚医学教育研究所(IMERI)实验室进行的体外研究。采用超离心法从ASC中分离外泌体。外泌体基于其颗粒大小、形态以及CD63和CD81表达的纯度进行了表征。我们能够通过差速超速离心从ASC中分离出无菌外泌体。外泌体的平均大小为88.7 nm±40 nm SD,并显示CD63和CD81的表达。使用超速离心法从ASC中成功分离出外泌体,并应按照MISEV标准进行表征,以满足作为再生剂的治疗效果和安全性问题。
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4 weeks
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