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Automated microinjection of cell-polymer suspensions in 3D ECM scaffolds for high-throughput quantitative cancer invasion screens.

机译:自动显微注射3D ECM支架中的细胞聚合物悬液,用于高通量定量癌症浸润筛查。

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摘要

Cell spheroids (CS) embedded in 3D extracellular matrix (ECM) serve as in vitro mimics for multicellular structures in vivo. Such cultures, started either from spontaneous cell aggregates or single cells dispersed in a gel are time consuming, applicable to restricted cell types only, prone to high variation, and do not allow CS formation with defined spatial distribution required for high-throughput imaging. Here, we describe a method where cell-polymer suspensions are microinjected as droplets into collagen gels and CS formation occurs within hours for a broad range of cell types. We have automated this method to produce CS arrays in fixed patterns with defined x-y-z spatial coordinates in 96 well plates and applied automated imaging and image analysis algorithms. Low intra- and inter-well variation of initial CS size and CS expansion indicates excellent reproducibility. Distinct cell migration patterns, including cohesive strand-like - and individual cell migration can be visualized and manipulated. A proof-of-principle chemical screen is performed identifying compounds that affect cancer cell invasion/migration. Finally, we demonstrate applicability to freshly isolated mouse breast and human sarcoma biopsy material - indicating potential for development of personalized cancer treatment strategies.
机译:嵌入3D细胞外基质(ECM)中的细胞球体(CS)用作体内多细胞结构的体外模拟物。从自发细胞聚集体或分散在凝胶中的单个细胞开始的此类培养非常耗时,仅适用于受限制的细胞类型,容易发生高变异,并且不允许形成具有高通量成像所需的确定空间分布的CS。在这里,我们描述了一种方法,其中将细胞聚合物悬浮液以液滴的形式微注射到胶原蛋白凝胶中,并在数小时内针对多种细胞类型发生CS形成。我们已经自动化了该方法,以在96孔板中以固定的模式生成具有定义的x-y-z空间坐标的CS阵列,并应用了自动成像和图像分析算法。初始CS尺寸和CS扩展的孔内和孔间变化小,表明可重复性极好。不同的细胞迁移模式(包括粘性链状)和单个细胞迁移可以可视化和操纵。进行原理证明化学筛选,以鉴定影响癌细胞侵袭/迁移的化合物。最后,我们证明了对新鲜分离的小鼠乳房和人肉瘤活检材料的适用性-表明了开发个性化癌症治疗策略的潜力。

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