Basic structure and cytocompatibility of giant membrane vesicles derived from paraformaldehyde-exposed human cells.
Basic structure and cytocompatibility of giant membrane vesicles derived from paraformaldehyde-exposed human cells.
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来自多聚甲醛暴露的人体细胞的巨膜囊泡的基本结构和细胞相容性。
DOI：
10.1093/jb/mvab144
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发表时间：
2021
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;Yuki Tanoue;Hesham Nasser;Takaichi Fukuda;Terumasa Ikeda;Hisato Saitoh
将培养的哺乳动物细胞暴露于多聚甲醛（PFA）是诱导膜泡的有效方法，随后膜泡从细胞皮层脱离以在细胞外空间产生巨大的膜泡。虽然PFA诱导的巨囊泡在细胞膜动力学领域引起了极大的兴趣，但它们的生化成分和细胞相容性在很大程度上仍然未知。在本报告中，我们在无金属缓冲液条件下将人宫颈癌HeLa细胞暴露于PFA以产生巨大的囊泡。我们分析了纯化的PFA诱导的巨囊泡的组成和结构。共培养PFA诱导的巨囊泡与指数生长的HeLa细胞导致大量的巨囊泡与细胞表面对接，似乎没有细胞毒性。有趣的是，我们发现用肽-N-糖苷酶和神经氨酸酶预处理HeLa细胞有效地促进细胞摄取囊泡内的成分。结果揭示了PFA对细胞膜作用的进一步细节，并为研究PFA诱导的巨囊泡与培养细胞之间的相互作用提供了见解。
Exposure of cultured mammalian cells to paraformaldehyde (PFA) is an effective approach to induce membrane blebs, which is followed by their detachment from the cellular cortex to yield giant membrane vesicles in extracellular spaces. Although PFA-induced giant vesicles have attracted significant interest in the field of cell membrane dynamics, their biochemical components and cytocompatibility remain largely unknown. In this report, we exposed human cervical cancer HeLa cells to PFA under metal-free buffer conditions to produce giant vesicles. We analyzed the components and structure of the purified PFA-induced giant vesicles. Co-culturing PFA-induced giant vesicles with exponentially growing HeLa cells resulted in docking of a significant number of the giant vesicles to the cell surface with seemingly no cytotoxicity. Intriguingly, we found that pre-treatment of HeLa cells with peptide-N-glycosidase and neuraminidase was effective in facilitating cellular uptake of constituents residing inside the vesicles. The results revealed further details about the effect of PFA on cell membranes and provide insights for studying the interaction between PFA-induced giant vesicles and cultured cells.