Patterning Mammalian Cells for Modeling Three Types of Naturally Occurring Cell-Cell Interactions
Patterning Mammalian Cells for Modeling Three Types of Naturally Occurring Cell-Cell Interactions
复制标题
哺乳动物细胞图案化以模拟三种自然发生的细胞间相互作用
DOI：
10.1002/anie.200902708
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发表时间：
2009-01-01
影响因子：
16.6
通讯作者：
Jiang, Xingyu
中科院分区：
文献类型：
--
作者：
Chen, Zhenling;Li, Yong;Jiang, Xingyu
在此，我们报告了一种在体外模拟三种类型的细胞-细胞相互作用的方法，通过微流控通道对表面进行选择性修饰。不同类型细胞之间的体内相互作用包括以下几种类型：1)两种类型细胞之间的相互作用，这些相互作用都是固定的，并且局限于孤立的区域（为简单起见，我们称这种相互作用为“I型”），例如卵巢发育过程中上皮细胞和成纤维细胞，上皮细胞和极性细胞；[1] 2)介于一种不活动的细胞类型和另一种自由活动的细胞类型（“ii型”）之间的细胞，如神经系统发育期间和各种神经退行性疾病中的胶质细胞和神经元；[3]和3)介于两种或两种以上自由移动的细胞之间（“III型”），例如肝脏中的肝细胞和成纤维细胞生物体的发育、神经网络的建立、肿瘤的形成，都涉及到上述所有不同类型细胞之间的相互作用。然而，没有一种现有的技术可以动态地控制所有这些行为，并以微米级的精度模拟所有三种类型的细胞-细胞相互作用尽管我们有大量的方法可以在固体表面上对生物分子（如蛋白质）进行建模，但哺乳动物细胞与固体表面之间相互作用的两个特点使得很难对多种类型的细胞进行建模：1)大多数贴壁哺乳动物细胞在体外培养时随机移动和分散；2)新播种的细胞往往会挤进最紧密的单层细胞中，并粘附在细胞表面，导致细胞群体不均匀。这些特征使得多种类型细胞的图像化，换句话说，控制哺乳动物细胞的粘附和迁移以允许所有三种类型的细胞-细胞相互作用的能力，与多种类型蛋白质的图像化相比仍然相当困难。几个研究小组已经报告了绘制多种类型细胞的技术；其中一些方法允许将两种或两种以上类型的细胞限制在表面上的特定位置（模拟类型I），[5,6]以及这些不同类型的细胞相互移动（模拟类型iii）。[5,7]然而，据我们所知，尽管II型细胞-细胞相互作用是正常和病理生理学中经常遇到的一种重要的细胞-细胞相互作用，但还没有发表的研究证明了II型细胞-细胞相互作用的模拟。[2,8]在此，我们提出了一种实现所有三种类型细胞-细胞相互作用的新方法：“不释放”（多种类型细胞的限制和分离，类型I），“部分释放”（一种细胞类型的选择性释放以自由运动，同时保持另一种细胞类型的限制，类型ii），以及“完全释放”（所有类型细胞的完全释放，类型III）多种类型细胞（图1）。我们的方法使用烷硫醇[(HS (CH2) 11 (OCH2CH2) 6OH，缩写为EG6][9]来生成抵抗细胞粘附（“惰性”区域）的自组装单层（sam），以及从溶液中物理吸附的细胞外基质蛋白纤维连接蛋白（FN）来生成促进细胞粘附的表面（“允许”区域）；我们使用微通道来创建具有不同表面特性（允许或惰性）的区域，并将不同类型的细胞运输到同一表面的指定位置。我们的策略比以前报道的方法简单得多。我们坚信这一策略将在细胞生物学的基础研究中得到广泛应用。我们首先演示……的方法。
Herein we report a method for the simulation of three types of cell–cell interactions in vitro on the same substrate by using selective modification of the surface by microfluidic channels. In vivo interactions between different types of cells include the following types: 1) those between two types of cells that are both immobilized and confined to isolated areas (for brevity, we call this kind of interaction “type I”), for example, epithelial cells and fibroblasts, and epithelial cells and polar cells during ovarian development;[1] 2) those between one cell type that is immobile and another that moves freely (“typeII”), such as glial cells and neurons during the development of the nervous system and in various neurodegenerative disorders;[2] and 3) those between two or more types of cells that are both moving freely (“type III”), for example, hepatocytes and fibroblasts in the liver.[3] The development of an organism, the establishment of neural networks, and the formation of tumors involve all aforementioned types of interactions between different types of cells. None of the existing techniques can, however, dynamically control all of these behaviors and model all three types of cell–cell interactions with micrometer-scale precision.[4] Even though we have a large number of available methods to pattern biological molecules (such as proteins) on solid surfaces, two characteristics of the interactions between mammalian cells and solid surfaces make it difficult to pattern multiple types of cells: 1) Most adherent mammalian cells, when cultured in vitro, randomly move and scatter around; and 2) newly seeded cells tend to squeeze into even the most tightly formed monolayers of cells and adhere to the surface, resulting in nonhomogeneous populations of cells in patterns. These features make patterning of multiple types of cells, in other words, the ability to control the adhesion and migration of mammalian cells to allow all three types of cell–cell interactions, still quite difficult in comparison to the patterning of multiple types of proteins. Several research groups have reported techniques for patterning multiple types of cells; some of these methods allow the confinement of two or more types of cells to specific locations on surfaces (to simulate type I),[5, 6] as well as the movement of these different types of cells toward each other (to simulate typeIII).[5, 7] To our knowledge, however, no published work demonstrates the simulation of type II cell–cell interactions though it is an important type of cell–cell interaction often encountered in normal and pathological physiology.[2, 8]Herein, we present a new method that achieves all three types of cell–cell interactions:“no release”(the confinement and segregation of multiple types of cells, type I),“partial release”(the selective release of one cell type for free motility while keeping the other cell type confined, typeII), and “complete release”(the complete release of all types of cells, type III) of multiple types of cells (Figure 1). Our method uses an alkanethiol [(HS (CH2) 11 (OCH2CH2) 6OH, abbreviated as EG6][9] to generate self-assembled monolayers (SAMs) that resist cell adhesion (“inert” areas), and the extracellular matrix protein fibronectin (FN) physically adsorbed from solution to generate surfaces that promote cell adhesion (“permissive” areas); we employ microchannels both to create areas with different surface properties (either permissive or inert) and to transport different types of cells to designated locations on the same surface. Our strategy is dramatically simpler than previously reported methods. We strongly believe that this strategy will find wide application in fundamental research in cell biology. We first demonstrate the approach to …