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首页> 外文期刊>Biomaterials Science >Generation of 3D cellular spheroids using DNA modified cell receptors and programmable DNA interactions
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Generation of 3D cellular spheroids using DNA modified cell receptors and programmable DNA interactions

机译：使用 DNA 修饰的细胞受体和可编程的 DNA 相互作用生成 3D 细胞球体

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3D culture is known to provide more faithful tissue models than 2D culture, and thus it is a valuable tool for in vitro evaluation of biological models. However, many cell lines are unable to form desired 3D spheroids by traditional methods because the naturally occurring cell-cell adhesion is too weak. Here, we present a method to produce 3D cell spheroids by using DNA-mediated assembly. We first demonstrate an Affinity Mediated Photoconjugation Approach (AMCP) to covalently modify cell receptors with affibody-streptavidin fusion proteins, where the affibody chemically crosslinks to cell expressed EGFR and the streptavidin is used to attach DNA strands. The DNA conjugated cells were then mixed with complementary DNA 'linker strands' to impart cell-cell interactions. When incubated in wells coated with non-adhesive polymers, cells formed dense spherical aggregates larger than 500 microns in diameter. Each of these studies was carried out using human breast cancer cells (MBA-MB-468), aneuploid human keratinocytes (HaCaT), and human colon cancer cells (Caco-2). Without either DNA on the cells or in solution as linkers, no cell spheroids were observed. After 96 h of incubation, the cultured DNA assembled spheroids were found to be mechanically stable enough to be handled easily for further analysis and confocal imaging. The findings suggest that the proposed DNA assembly method can be considered as an attractive strategy for assembling cells into stable spheroids.
机译：众所周知，3D 培养比 2D 培养提供更忠实的组织模型，因此它是体外评估生物模型的宝贵工具。然而，许多细胞系无法通过传统方法形成所需的 3D 球状体，因为天然存在的细胞间粘附太弱。在这里，我们提出了一种通过使用 DNA 介导的组装来生产 3D 细胞球体的方法。我们首先展示了一种亲和介导的光偶联方法（AMCP），用亲和体-链霉亲和素融合蛋白共价修饰细胞受体，其中亲和体与细胞表达的EGFR化学交联，链霉亲和素用于连接DNA链。然后将DNA偶联的细胞与互补的DNA“连接链”混合，以传递细胞间相互作用。当在涂有非粘性聚合物的孔中孵育时，细胞形成直径大于 500 微米的致密球形聚集体。这些研究中的每一项都是使用人乳腺癌细胞 （MBA-MB-468）、非整倍体人角质形成细胞 （HaCaT） 和人结肠癌细胞 （Caco-2） 进行的。在细胞上或溶液中均未发现DNA作为接头，因此未观察到细胞球状体。孵育 96 小时后，发现培养的 DNA 组装球体具有足够的机械稳定性，易于处理以进行进一步分析和共聚焦成像。研究结果表明，所提出的DNA组装方法可以被认为是将细胞组装成稳定球状体的有吸引力的策略。

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