Construction of an In Vitro Model of a Living Cellular System
Construction of an In Vitro Model of a Living Cellular System
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活细胞系统体外模型的构建
DOI：
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发表时间：
2011
期刊：
影响因子：
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通讯作者：
K. Takiguchi
中科院分区：
文献类型：
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作者：
K. Yoshikawa;S. Nomura;K. Tsumoto;K. Takiguchi
使用与活细胞完全相容的材料可能是生产人工细胞模型的最合理方案。在这篇文章中，我们提出了一些最新的研究结果，对人工细胞的建设，并解释方法模拟几个重要的特定功能的真实的细胞在人工脂质体。我们讨论了当前实验方法的几个方面，从巨囊泡（GVs）的优化构建到复杂囊泡分子系统的实现。讨论了GVs内DNA和酶的截留，以及囊泡内水溶性蛋白质的合成。重点介绍了在囊泡膜上合成膜溶性蛋白的新方法。连接蛋白43（Cx43）的情况下，含有脂质体的详细描述，以及延伸的蛋白脂质体技术的巨大的脂质体，适合直接显微镜成像。目前在体外合成方法的细胞形态学的研究中的应用进行了讨论，通过参考部分重建的细胞骨架内GVs。在我们最近的研究中，G-actin和BBMI同时包埋在脂质体中，通过自然溶胀的方法，导致有效的GV转化。我们的结论与一些一般性的考虑未来的工作，细胞模型使用囊泡。
The use of materials that are fully compatible with living cells may be the most reasonable plan for producing artificial models of cells. In this contribution, we present the results of some recent studies toward the construction of artificial cells, and explain methods for emulating several important specific functions of real cells in artificial liposomes. We discuss several aspects of the current experimental approaches, from the optimal construction of giant vesicles (GVs) to the realization of complex vesicle-based molecular system. The entrapment of DNA and enzymes inside GVs are discussed, as well as the synthesis of water soluble proteins inside vesicles. Emphasis is given on the new approaches to synthesize membrane-soluble proteins on the vesicle membrane. The case of connexin 43 (Cx43)-containing liposomes is described in details, as well as the extension of the proteoliposome technology to giant lipsomes, suitable for direct microscopy imaging. Application of the current in vitro synthetic approaches to the study of cell morphology is also discussed, by referring to the partial reconstitution of cytoskeleton inside GVs. In our recent study, G-actin and BBMI were simultaneously entrapped inside liposomes, through the method of natural swelling, resulting in efficient GV transformation. We conclude with some general consideration about the future work on cell models using vesicles.