Natsume Yuno, Wen Hsin-I, Zhu Tong, Itoh Kazumi, Sheng Li, Kurihara Kensuke
Department of Mathematical and Physical Sciences, Faculty of Science, Japan Women's University;
Department of Bioorganization Research, Okazaki Institute for Integrative Bioscience, National Institutes of Natural Sciences.
J Vis Exp. 2017 Jan 24(119):55282. doi: 10.3791/55282.
The constructive biology and the synthetic biology approach to creating artificial life involve the bottom-up assembly of biological or nonbiological materials. Such approaches have received considerable attention in research on the boundary between living and nonliving matter and have been used to construct artificial cells over the past two decades. In particular, Giant Vesicles (GVs) have often been used as artificial cell membranes. In this paper, we describe the preparation of GVs encapsulating highly packed microspheres as a model of cells containing highly condensed biomolecules. The GVs were prepared by means of a simple water-in-oil emulsion centrifugation method. Specifically, a homogenizer was used to emulsify an aqueous solution containing the materials to be encapsulated and an oil containing dissolved phospholipids, and the resulting emulsion was layered carefully on the surface of another aqueous solution. The layered system was then centrifuged to generate the GVs. This powerful method was used to encapsulate materials ranging from small molecules to microspheres.
构建生物学和合成生物学中创造人工生命的方法涉及从生物或非生物材料自下而上进行组装。此类方法在关于生命与非生命物质边界的研究中受到了广泛关注，并且在过去二十年里一直被用于构建人工细胞。特别是，巨型囊泡（GVs）常被用作人工细胞膜。在本文中，我们描述了制备包裹高度密集微球的巨型囊泡，以此作为包含高度浓缩生物分子的细胞模型。巨型囊泡通过一种简单的油包水乳液离心法制备。具体而言，使用均质器将含有待包裹材料的水溶液与含有溶解磷脂的油相乳化，然后将所得乳液小心地铺在另一种水溶液表面。接着对分层体系进行离心以生成巨型囊泡。这种强大的方法被用于包裹从小分子到微球等各种材料。
