Trehalose prevents aggregation of exosomes and cryodamage.
Trehalose prevents aggregation of exosomes and cryodamage.
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DOI：
10.1038/srep36162
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发表时间：
2016-11-08
影响因子：
4.6
通讯作者：
Bach JM
中科院分区：
文献类型：
--
作者：
Bosch S;de Beaurepaire L;Allard M;Mosser M;Heichette C;Chrétien D;Jegou D;Bach JM
外泌体是细胞间通讯的重要介质。它们由许多细胞类型释放，将蛋白质、脂质和核酸转运到远处的受体细胞，并促进重要的病理生理过程。目前基于差速离心方案的标准外泌体分离方法往往会引起高浓度悬浮液中颗粒的聚集，而外泌体的冷冻可能会导致损伤和不一致的生物活性。海藻糖是一种天然、无毒的糖，在食品和药品工业中广泛用作蛋白质稳定剂和冷冻保护剂。在此，我们报告在胰腺 β 细胞外泌体样囊泡分离和储存缓冲液中添加 25 mM 海藻糖可以缩小粒径分布，并增加每微克蛋白质的单个颗粒数量。重复冻融循环会导致储存在 PBS 中的外泌体样囊泡的颗粒浓度和尺寸分布宽度增加，但在 PBS 25mM 海藻糖中则不会。通过冷冻电子断层扫描在 PBS 和海藻糖样品中未观察到裂解或不完整囊泡的迹象。在巨噬细胞免疫测定中，海藻糖中的 β 细胞胞外囊泡显示出一致较高的 TNF-α 细胞因子分泌刺激指数，表明生物活性的保存得到改善。添加海藻糖可能是外泌体研究和下游应用领域实验标准化的一种有吸引力的方法。
Exosomes are important mediators in intercellular communication. Released by many cell types, they transport proteins, lipids, and nucleic acids to distant recipient cells and contribute to important physiopathological processes. Standard current exosome isolation methods based on differential centrifugation protocols tend to induce aggregation of particles in highly concentrated suspensions and freezing of exosomes can induce damage and inconsistent biological activity. Trehalose is a natural, non-toxic sugar widely used as a protein stabilizer and cryoprotectant by the food and drug industry. Here we report that addition of 25 mM trehalose to pancreatic beta-cell exosome-like vesicle isolation and storage buffer narrows the particle size distribution and increases the number of individual particles per microgram of protein. Repeated freeze-thaw cycles induce an increase in particle concentration and in the width of the size distribution for exosome-like vesicles stored in PBS, but not in PBS 25 mM trehalose. No signs of lysis or incomplete vesicles were observed by cryo-electron tomography in PBS and trehalose samples. In macrophage immune assays, beta-cell extracellular vesicles in trehalose show consistently higher TNF-alpha cytokine secretion stimulation indexes suggesting improved preservation of biological activity. The addition of trehalose might be an attractive means to standardize experiments in the field of exosome research and downstream applications.
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影响因子：
7.4
作者：
Echigo R;Shimohata N;Karatsu K;Yano F;Kayasuga-Kariya Y;Fujisawa A;Ohto T;Kita Y;Nakamura M;Suzuki S;Mochizuki M;Shimizu T;Chung UI;Sasaki N
通讯作者：
Sasaki N
DOI：
10.1016/j.bbrc.2008.06.094
发表时间：
2008-09-12
影响因子：
3.1
作者：
Beranger, Florence;Crozet, Carole;Lehmann, Sylvain
通讯作者：
Lehmann, Sylvain
影响因子：
4.6
作者：
Katenz, Ekaterina;Vondran, Florian Wolfgang Rudolf;Sauer, Igor Maximilian
通讯作者：
Sauer, Igor Maximilian
影响因子：
3
作者：
Kremer, JR;Mastronarde, DN;McIntosh, JR
通讯作者：
McIntosh, JR
影响因子：
2.5
作者：
Ikeda, Masaki;Bando, Toru;Date, Hiroshi
通讯作者：
Date, Hiroshi