Structurally-reduced cells and cell-derived structures are powerful tools for membrane studies. Using this approach, we probed whether a cell, without its nucleus and cytoplasm, is still capable of undergoing CD4-mediated membrane fusion. For this, we needed a cell-derived structure, akin to a giant liposome functionalised with CD4 and chemokine receptors. We present a method for the simultaneous removal of cytoplasmic and nuclear material from cells presenting CD4, CCR5, and CXCR4, using Colcemid treatment followed by hypotonic cytolysis, and then enriched using preparative flow cytometry. We show that the resultant cell membrane remains intact, retains presentation of CD4, CCR5, and CXCR4, and is still capable of CD4-mediated membrane fusion with a target cell. Finally, we detail how this protocol was developed, as well as how such samples should be handled for storage and assays. We envision the use of such systems for host-pathogen interaction studies, and the development of targeted delivery vehicles.

结构减少的细胞和细胞衍生结构是膜研究的有力工具。使用这种方法，我们探测了没有细胞核和细胞质的细胞是否仍然能够进行 CD4 介导的膜融合。为此，我们需要一种细胞衍生结构，类似于用 CD4 和趋化因子受体功能化的巨型脂质体。我们提出了一种从呈递 CD4、CCR5 和 CXCR4 的细胞中同时去除细胞质和核材料的方法，使用秋水仙碱处理，然后进行低渗细胞溶解，然后使用制备型流式细胞术进行富集。我们表明由此产生的细胞膜保持完整，保留了 CD4、CCR5 和 CXCR4 的呈现，并且仍然能够与目标细胞进行 CD4 介导的膜融合。最后，我们详细介绍了这个协议是如何开发的，以及应如何处理此类样品以进行储存和分析。我们设想使用这样的系统进行宿主-病原体相互作用研究，以及开发有针对性的运载工具。