3D printing of microscopic bacterial communities
3D printing of microscopic bacterial communities
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DOI：
10.1073/pnas.1309729110
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发表时间：
2013-11-12
影响因子：
11.1
通讯作者：
Shear, Jason B.
中科院分区：
文献类型：
--
作者：
Connell, Jodi L.;Ritschdorff, Eric T.;Shear, Jason B.
细菌通过短距离的物理和化学信号进行交流，已知的相互作用介导群体感应、产孢和其他适应性表型。尽管大多数体外研究检查了在大群体中平均的细菌特性，但细菌适合度和致病性的关键分子决定因素（例如氧气、群体感应信号）的水平可能在微米尺度上在被认为在疾病传播中起关键作用的小而密集的细胞聚集体内发生变化。要详细了解细胞-细胞相互作用如何在自然、复杂环境中促进致病性，就需要在构建更相关的细胞模型以评估细菌表型方面达到一个新的控制水平。在这里，我们描述了一种微观三维（3D）打印策略，使多种细菌群体能够在本质上任何3D几何结构中组织起来，包括相邻的、嵌套的和自由浮动的菌落。在这种基于激光的光刻技术中，通过多肽分子的焦点交联，在悬浮在明胶中的选定细菌周围形成微观容器。在去除多余的试剂后，被困的细菌被定位在由交联明胶形成的密封腔内。明胶是一种高度多孔的材料，支持全封闭细胞群的快速生长，并容易传播许多生物活性物质，包括多肽、抗生素和群体感应信号。使用这种方法，我们发现皮升体积的金黄色葡萄球菌聚集在铜绿假单胞菌组成的外壳内可以显示出对β -内酰胺类抗生素的实质性耐药性。
Bacteria communicate via short-range physical and chemical signals, interactions known to mediate quorum sensing, sporulation, and other adaptive phenotypes. Although most in vitro studies examine bacterial properties averaged over large populations, the levels of key molecular determinants of bacterial fitness and pathogenicity (e. g., oxygen, quorum-sensing signals) may vary over micrometer scales within small, dense cellular aggregates believed to play key roles in disease transmission. A detailed understanding of how cell-cell interactions contribute to pathogenicity in natural, complex environments will require a new level of control in constructing more relevant cellular models for assessing bacterial phenotypes. Here, we describe a microscopic three-dimensional (3D) printing strategy that enables multiple populations of bacteria to be organized within essentially any 3D geometry, including adjacent, nested, and free-floating colonies. In this laser-based lithographic technique, microscopic containers are formed around selected bacteria suspended in gelatin via focal cross-linking of polypeptide molecules. After excess reagent is removed, trapped bacteria are localized within sealed cavities formed by the crosslinked gelatin, a highly porous material that supports rapid growth of fully enclosed cellular populations and readily transmits numerous biologically active species, including polypeptides, antibiotics, and quorum-sensing signals. Using this approach, we show that a picoliter-volume aggregate of Staphylococcus aureus can display substantial resistance to beta-lactam antibiotics by enclosure within a shell composed of Pseudomonas aeruginosa.