Chemica manipulation to facilitate membrane blebbing and vesicle shedding in the cellular cortex.
Chemica manipulation to facilitate membrane blebbing and vesicle shedding in the cellular cortex.
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化学操作促进细胞皮层中的膜起泡和囊泡脱落。
DOI：
10.1007/s10529-020-02848-7
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发表时间：
2020
期刊：
影响因子：
--
通讯作者：
Hisato Saitoh
中科院分区：
文献类型：
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作者：
Saya Okada;Yuta Fukai;Fumiya Yoshimoto;Hisato Saitoh
目的研究主要集中在细胞皮层生理性产生的膜泡，而化学物质诱导的人工膜泡的研究较少。结果我们发现HeLa人宫颈癌细胞暴露于多聚甲醛（PFA）中，然后在磷酸盐缓冲盐水（PBS）中孵育，有效地诱导了细胞皮层上的大膜泡。有趣的是，将经pfa处理的细胞连续暴露于含有二甲亚砜（DMSO）的PBS中，促进了细胞皮层的气泡脱落，在上清中产生大量的大细胞外囊泡，这适用于评估化合物和蛋白质作为膜影响者的潜力。PFA和DMSO对其他人类、小鼠和鱼类细胞的皮层也有类似的影响。结论利用化学物质促进膜起泡和囊泡脱落的方法，可用于多种细胞类型的膜动力学操作和囊泡激发技术的开发。
ObjectivesMost attention has been focused on physiologically generated membrane blebs on the cellular cortex, whereas artificial membrane blebs induced by chemicals are studied to a lesser extent.ResultsWe found that exposure of HeLa human cervical cancer cells to paraformaldehyde (PFA), followed by incubation in phosphate-buffered saline (PBS) efficiently induced large membrane blebs on the cellular cortex. Intriguingly, sequential exposure of the PFA-treated cells to PBS containing dimethyl sulfoxide (DMSO) facilitated shedding of the blebs from the cellular cortex, yielding a high quantity of large extracellular vesicles in the supernatant, which was applicable to assess the potentials of compounds and proteins as membrane influencers. Similar effects of PFA and DMSO were detected on the cellular cortex of other human, mouse, and fish cells.ConclusionsOur procedure to facilitate membrane blebbing and vesicle shedding by chemicals may be practical for the manipulation of membrane dynamics and the development of vesicle-inspired technologies using a wide variety of cell types.