High throughput scaffoldless 3D tissue culture kit

传统上，细胞以平面二维（2D）结构的形式生长，并假设它们近似于组织、器官和肿瘤的三维行为。由于二维细胞结构不能准确地模拟生物体内复杂的三维排列之间的细胞-细胞和细胞-基质相互作用，研究人员已经转向使用三维（3D）细胞结构来发现和筛选药物。因此，生产3D结构的新方法和设备已经开发并商业化，但都面临限制。它们非常耗时，需要紫外线照射，对细胞有毒，或者相对昂贵且操作复杂。这限制了它们在学术界和工业界的采用和广泛使用。我们的方法是使用反磁泳术来操纵细胞以产生3D细胞结构，这是一种强大而有前途的替代方法。它克服了现有方法的缺点，作为一种无标签、无支架和可扩展的技术，可以将细胞快速打印成不同大小的球体或环。我们将细胞悬浮在顺磁溶液中，并将悬浮种子放入微孔中，微孔随后放置在磁体阵列上。由于外加磁场的梯度，抗磁性细胞在三小时内迁移并形成聚集体。我们的现场辅助技术不需要物理插入或矩阵，因此3D细胞结构可以独立生长。整个过程可以实现自动化和规模化，从而在多孔板上生成丰富的3D结构。我们已经在麦克马斯特大学的实验室演示了概念验证。该技术将进一步发展，以便系统原型可以用于确定的最终用户的测试。一旦我们的技术得到验证，我们将与感兴趣的行业合作伙伴一起研究许可选项。

Cells have been traditionally grown in the form of planar two-dimensional (2D) constructs with the assumption that they approximate the 3D behaviour of tissues, organs and tumours. Since 2D cell structures cannot accurately mimic cell-cell and cell-matrix interactions among the complex 3D arrangements within living organisms, researchers have pivoted to using three-dimensional (3D) cellular structures to discover and screen drugs. Hence, new methods and devices to produce 3D structures have been developed and commercialized, but all face limitations. They are inordinately time-consuming, require UV light exposure, are toxic towards cells, or are relatively expensive and complex to operate. This has restricted their adoption and wide use across academia and industry. Our method to manipulate cells using diamagnetophoresis to produce 3D cellular structures is a robust and promising alternative. It overcomes the shortcomings of available methods as a label-free, scaffold-free and scalable technique that prints cells rapidly into spheroids or rings of different sizes. We suspend cells in a paramagnetic solution and seed the suspension into microwells that are subsequently placed on a magnet array. Due to the applied magnetic field gradient, diamagnetic cells migrate and form aggregates within three hours. Our field-assisted technique does not require physical inserts or a matrix so that the 3D cellular structures can grow independently. The entire process can be automated and scaled up to produce abundant 3D structures on a multiwell plate. We have demonstrated proof of concept in our laboratory at McMaster University. The technology will be developed further so that system prototypes can be used for testing by identified end-users. Once our technology is validated, we will examine licensing options with interested industry partners.