Name: constant pressure-controlled extrusion method for the preparation of nano-sized lipid vesicles
Uploaded: 2012-06-22T13:00:00
Description: Watch this Scientific Journal Video about Constant Pressure-controlled Extrusion Method for the Preparation of Nano-sized Lipid Vesicles at JoVE.com

这项工作得到了霍华德·休斯医学研究所（HHMI）协同创新奖。林支持信令和细胞调节研究所的T32 GM008759卫生培训资助和NIH的露丝L. Kirschstein前博士后研究员（CA165349-01）。我们要感谢他们的宝贵意见教授迈克尔·斯托厄尔（铜博尔德），教授道格拉斯里斯和罗布·菲利普斯教授（加州理工学院）。

1。脂质体的制备<ol><li>检索与聚四氟乙烯内衬盖20 mL的玻璃小瓶。 </li><li>清理所有与氯仿的玻璃器皿和注射器使用前，以防止污染。 </li><li>转移100μL试剂级氯仿的玻璃小瓶，用250μL气密玻璃注射器。 </li><li>加入30μL试剂级甲醇相同的玻璃小瓶，用100μL气密玻璃注射器。 </li><li>准备一个2毫米7:1.5:1.5卵磷脂（POPC）：磷脂（教皇）：胆固醇脂解，转移216μL10毫克/ 10毫克/毫升教皇和20μL11毫克/毫升，...

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我们使用的Avestin Liposofast LF-50挤出机，并演示了如何小型，合成脂质体通过压力控制系统的准备。重要的是要注意多室囊泡自发形成脂质体水化后，这可能会导致更小的纳米粒子的生产。这些小的多室囊泡，将不可避免地流过较大的聚碳酸酯膜孔径，导致异质性的解决方案由一个大型的过滤孔的单层囊泡。因此，建议，以减少氮流量压力，当使用大孔径直径为400纳米以上所述毛孔（即25磅）为了?...

<table border="1"><tbody><tr><td> 试剂名称 </td><td> 公司 </td><td> 目录编号 </td><td> 评论 </td></tr><tr><td>氯仿</td><td> Sigma-Aldrich公司</td><td> 02432-25ML </td><td> 95％的稳定剂</td></tr><tr><td>高品位的甲醇</td><td> Sigma-Aldrich公司</td><td> 179337-4L </td><td></td></tr><tr><td> liposofast LF-50挤出机</td><td> avestin，公司</td><td></td><td></td></tr><tr><td>磷脂</td><td>阿凡提极性脂</td><td></td><td></td></tr><tr><td>聚碳酸酯毛孔</td><td> avestin，公司</td><td></td><td>直径25毫米</td></tr><tr><td>排水光盘体育</td><td> avestin，公司</td><td> 230600 </td><td>直径25毫米</td></tr></tbody></table>

constant pressure-controlled extrusion method for the preparation of nano-sized lipid vesicles

脂质体是人为编制的模拟平台，研究蛋白质和脂质蛋白相互作用3，监测给药4,5，和4封装膜作为细胞被广泛使用的天然和合成磷脂组成的囊泡。磷脂自然弯曲的脂质双层，本身区别于胶束。6脂质体是双层，即大（LUVs）的单层囊泡，小囊泡（越野车）单层和多室囊泡（MLV更具）的大小和数量由传统分类。特别是，编制各种大小均匀脂质体是重要的研究膜的曲率，在细胞信号传递中扮演了至关重要的作用，内和胞吐作用，膜融合，蛋白质贩运8。几组分析蛋白质如何被用来调节过程，涉及膜的曲率，从而准备脂质体直径&lt;100 - 400纳米研究其对细胞功能的行为3。其他专注于脂质体药物包封，学习车辆进行并交付利息9药物的脂质体。可以实现药物包封在脂质体形成9报告。我们的挤压步骤应该没有影响，原因有两个封装的药物，即（1）药物包封应事先取得到了这一步，（2）脂质体应保留其天然生物物理稳定性，安全携带药物在水溶液中的核心。这些研究的目标，进一步表明需要优化的方法来设计稳定的亚微米级的脂质囊泡。 尽管如此，目前的脂质体的制备技术（10超声，冷冻和解冻10，沉淀）不容许高度的一致性和高效率10,5，这限制了生物物理曲面（即直径小于100纳米）的脂质体的制备研究1 emergi会吴膜曲率传感领域。在此，我们提出了各种生物相关的脂质体健壮的制备方法。 使用气密注射器和聚碳酸酯薄膜10,5手动挤压是一种常见的做法，但经常被观察到的异质性时，使用孔径小于100纳米，由于由于手动施加压力的变异。我们采用恒压控制挤出设备，准备合成脂质体，其直径范围在30和400 nm之间。被用来量化脂质体的大小，在我们的协议与商业聚苯乙烯（PS）作为校准标准用于珠，动态光散射（DLS）10 11，电子显微镜和纳米粒子跟踪分析（NTA）的12。一个附近的线性相关就业孔径和实验测定脂质体之间，说明我们的压力控制脂质体的制备高保真会见HOD。此外，我们已经表明，这种脂质囊泡的制备方法是普遍适用的，独立于各种脂质体的大小。最后，我们在一个时间过程的研究还表明，这些准备脂质体是稳定的，长达16小时。下面演示了一个有代表性的纳米脂质体的制备协议。

Watch this Scientific Journal Video about Constant Pressure-controlled Extrusion Method for the Preparation of Nano-sized Lipid Vesicles at JoVE.com

Constant Pressure-controlled Extrusion Method for the Preparation of Nano-sized Lipid Vesicles

本协议描述了一个准备一个高度同质化的亚微米大小的脂质体的挤压方法。此方法使用脂质体的制备与控制氮流量压力控制系统。脂质准备<sup&gt; 1,2</sup&gt;，脂质体挤压，大小特征将在此呈现。

恒压控制的挤压方法制备纳米脂质体

Liposomes are artificially prepared vesicles consisting of natural and synthetic phospholipids that are widely used as a cell membrane mimicking platform ...

Research

JoVE Journal

Bioengineering

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改进方法对血脑屏障的人体细胞为主，接触模型的制备

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科研

生物工程

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对于I型胶原蛋白的制备皮肤方法的改进

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The Encapsulation of Cell-free Transcription and Translation Machinery in Vesicles for the Construction of Cellular Mimics

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As interest shifts from individual molecules to systems of molecules, an increasing number of laboratories have sought to build from the bottom up ...

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脂质双层囊泡代使用微流控喷射

Bottom-up synthetic biology presents a novel approach for investigating and reconstituting biochemical systems and, potentially, minimal organisms. This ...

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支持的脂双层云母制备的高分辨率光学显微成像

Supported lipid bilayers (SLBs) are widely used as a model for studying membrane properties (phase separation, clustering, dynamics) and its interaction ...

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生物膜制作的溶剂辅助脂质双分子层（SALB）方法

In order to mimic cell membranes, the supported lipid bilayer (SLB) is an attractive platform which enables in vitro investigation of membrane-related ...

Solid Lipid Nanoparticles (SLNs) for Intracellular Targeting Applications

Solid Lipid Nanoparticles SLNs for Intracellular Targeting Applications

固体脂质纳米粒（前哨淋巴结）细胞内定位的应用

Nanoparticle-based delivery vehicles have shown great promise for intracellular targeting applications, providing a mechanism to specifically alter ...

Film Extrusion of Crambe abyssinica/Wheat Gluten Blends

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薄膜挤出<em&gt;海甘蓝abyssinica</em&gt; /谷朊粉共混物

Crambe abyssinica is a plant with potential for use in industrial (non-food) plant oil production. The side stream from this oil production is a ...

Evaluation of the Storage Stability of Extracellular Vesicles

细胞外囊储存稳定性的评价

Extracellular vesicles (EVs) are promising targets in current research, to be used as drugs, drug-carriers, and biomarkers. For their clinical ...

Simultaneous Quantification of Selected Kynurenines Analyzed by Liquid Chromatography-Mass Spectrometry in Medium Collected from Cancer Cell Cultures

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The kynurenine pathway and the tryptophan catabolites called kynurenines have received increased attention for their involvement in immune regulation and ...