促进细胞膜起泡和小泡脱落的化学处理方法。
Chemical manipulations to facilitate membrane blebbing and vesicle shedding on the cellular cortex.
机构信息
Department of Biological Science, Graduate School of Science and Technology, Kumamoto University, Kumamoto, Japan.
Biology Course, Faculty of Science, Kumamoto University, Kumamoto, Japan.
出版信息
Biotechnol Lett. 2020 Jul;42(7):1137-1145. doi: 10.1007/s10529-020-02848-7. Epub 2020 Feb 28.
OBJECTIVES
Most 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.
RESULTS
We 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.
CONCLUSIONS
Our 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.
目的
大多数研究都集中在细胞皮层上由生理产生的膜泡上，而由化学物质诱导的人工膜泡则研究得较少。
结果
我们发现，将人宫颈癌细胞系（HeLa）暴露于多聚甲醛（paraformaldehyde，PFA）中，随后在磷酸盐缓冲盐水（phosphate-buffered saline，PBS）中孵育，可以有效地在细胞皮层上诱导大的膜泡。有趣的是，将经 PFA 处理的细胞依次暴露于含有二甲基亚砜（dimethyl sulfoxide，DMSO）的 PBS 中，促进了从细胞皮层上脱落的泡，在上清液中产生了大量的大细胞外囊泡，这可用于评估化合物和蛋白质作为膜影响物的潜力。在其他人类、小鼠和鱼类细胞的细胞皮层上也检测到了 PFA 和 DMSO 的类似作用。
结论
我们通过化学物质促进膜泡形成和囊泡脱落的方法，可能对使用各种细胞类型来操纵膜动力学和开发囊泡启发的技术具有实际意义。
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