A Novel Strategy to Increase the Yield of Exosomes (Extracellular Vesicles) for an Expansion of Basic Research
A Novel Strategy to Increase the Yield of Exosomes (Extracellular Vesicles) for an Expansion of Basic Research
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DOI：
10.1248/bpb.b17-00919
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发表时间：
2018-05-01
影响因子：
2
通讯作者：
Ishida, Tatsuhiro
中科院分区：
文献类型：
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作者：
Emam, Sherif E.;Ando, Hidenori;Ishida, Tatsuhiro
胞外体是一种微小的胞外小泡，通常是少量收获的。如此低的产率一直是扩大外切体分析的基础研究和在药物输送中应用的障碍。为了提高外切体产率，我们尝试在体外通过添加脂质体来刺激肿瘤细胞。将中性、阳离子裸露或聚乙二醇化的脂质体与四种不同的肿瘤细胞系孵育。评价脂质体制剂对母细胞或不同细胞收集的外切体的分泌和细胞摄取倾向的促进作用。中性和阳离子裸脂质体均以剂量依赖的方式促进外切体的分泌。液体阳离子脂质体的刺激性最强。令人惊讶的是，裸脂质体的聚乙二醇化减少了外切体的分泌。在液体阳离子脂质体存在下收获的外切体显示细胞摄取增加，但固体阳离子脂质体没有。我们的发现表明，脂质体的物理化学性质决定了它们是作为肿瘤细胞外体分泌的刺激剂还是抑制剂。脂质体刺激可能是提高外切体产量的一个有用的策略，尽管可能需要进一步的准备来提高外切体的纯度。此外，可以通过控制刺激性脂质体的物理化学性质来微调诱导外切体的生物学性质。
Exosomes are tiny extracellular vesicles that are usually harvested in small quantities. Such small yield has been an obstacle for the expansion of the basic research regarding exosome analysis and applications in drug delivery. To increase exosome yield, we attempted to stimulate tumor cells via the addition of liposomes in vitro. Neutral, cationic-bare or PEGylated liposomes were incubated with four different tumor cell lines. The stimulatory effect of liposomal formulations on exosome secretion and cellular uptake propensity of the collected exosome by mother cells or different cells was evaluated. Both neutral and cationic-bare liposomes enhanced exosome secretion in a dose-dependent manner. Fluid cationic liposomes provided the strongest stimulation. Surprisingly, the PEGylation of bare liposomes diminished exosome secretion. Exosomes harvested in the presence of fluid cationic liposomes showed increased cellular uptake, but solid cationic liposomes did not. Our findings indicate that the physicochemical properties of liposomes determine whether they will act as a stimulant or as a depressant on exosome secretion from tumor cells. Liposomal stimulation may be a useful strategy to increase exosome yield, although further preparation to increase the purity of exosomes may be needed. In addition, fine-tuning of the biological properties of induced exosomes could be achieved via controlling the physicochemical properties of the stimulant liposomes.