de Weerd Sander, Ruiter Emma A, Calicchia Eleonora, Portale Giuseppe, Schuringa Jan Jacob, Roos Wouter H, Salvati Anna
Nanomedicine and Drug Targeting, Groningen Research Institute of Pharmacy, University of Groningen, A. Deusinglaan 1, Groningen, 9713 AV, The Netherlands.
Molecular Biophysics, Zernike Institute, University of Groningen, Nijenborgh 4, Groningen, 9747 AG, The Netherlands.
Small Methods. 2024 Dec;8(12):e2400498. doi: 10.1002/smtd.202400498. Epub 2024 Oct 21.
Cell membrane nanoparticles have attracted increasing interest in nanomedicine because they allow to exploit the complexity of cell membrane interactions for drug delivery. Several methods are used to obtain plasma membrane to generate cell membrane nanoparticles. Here, an optimized method combining nitrogen cavitation in isotonic buffer and sucrose gradient fractionation is presented. The method allows to obtain cell membrane fractions of high purity from both suspension and adherent cells. Comparison with other common methods for membrane extraction, where mechanical lysis using cell homogenizers is performed in isotonic or hypotonic buffers, shows that the optimized procedure yields high purity membrane in a robust and reproducible way. Procedures to mix the purified membrane with synthetic lipids to obtain cell membrane liposomes (CMLs) are presented and indications on how to optimize these steps are provided. CMLs made using crude membrane isolates or the purified membrane fractions show different uptake by cells. The CMLs made with the optimized procedure and liposomes of the same composition but without cell membrane components are thoroughly characterized and compared for their size, zeta potential, bilayer and mechanical properties to confirm membrane protein inclusion in the CMLs. Cell uptake studies confirm that the inclusion of membrane components modifies liposome interactions with cells.
细胞膜纳米颗粒在纳米医学领域引起了越来越多的关注，因为它们能够利用细胞膜相互作用的复杂性来进行药物递送。有几种方法可用于获取质膜以制备细胞膜纳米颗粒。在此，我们介绍一种优化方法，该方法结合了等渗缓冲液中的氮空化和蔗糖梯度分级分离。该方法能够从悬浮细胞和贴壁细胞中获得高纯度的细胞膜组分。与其他常见的膜提取方法（即在等渗或低渗缓冲液中使用细胞匀浆器进行机械裂解）相比，结果表明该优化方法能够以稳健且可重复的方式获得高纯度的膜。文中介绍了将纯化的膜与合成脂质混合以获得细胞膜脂质体（CML）的步骤，并提供了如何优化这些步骤的建议。使用粗制膜分离物或纯化的膜组分制备的CML对细胞的摄取情况不同。对采用优化方法制备的CML以及具有相同组成但不含细胞膜成分的脂质体进行了全面表征，并比较了它们的大小、zeta电位、双层结构和机械性能，以确认CML中包含膜蛋白。细胞摄取研究证实，膜成分的加入改变了脂质体与细胞的相互作用。
