Advanced Micromachining of Concave Microwells for Long Term On-Chip Culture of Multicellular Tumor Spheroids
Advanced Micromachining of Concave Microwells for Long Term On-Chip Culture of Multicellular Tumor Spheroids
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DOI：
10.1021/am500367h
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发表时间：
2014-06-11
影响因子：
9.5
通讯作者：
Thierry, Benjamin
中科院分区：
文献类型：
--
作者：
Liu, Tianqing;Chien, Chia-Chi;Thierry, Benjamin
一种新的方法，先进的微加工的基础上，证明了制造凹微孔阵列形成高品质的多细胞肿瘤球体。使用最先进的CNC加工中心制备微制造模具，其含有尺寸范围从150 μ m到600 μ m的3D凸微柱阵列。在模具上加工的微柱的显微成像显示出戏剧性的3D形状的平滑、弯曲的微特征。琼脂糖微孔可以很容易地从金属模具复制。接种在原代微孔中的EMT-6肿瘤细胞在48 h内有效地沉积到凹形微孔的底部并形成多细胞球体。培养10天后获得了致密均匀的多细胞球体，证实了所提出的方法的适用性。为了促进长期球状体培养和可靠的芯片上药物测定，还从金属模具复制聚二甲基硅氧烷微孔。一种溶剂溶胀方法被调整和优化，以普朗尼克F127物理截留聚二甲基硅氧烷网络内的嵌段共聚物分子，从而提高长期的细胞结合阻力。均匀的多细胞球体有效地形成在凹微孔和芯片上的药物测定可以可靠地进行使用姜黄素作为模型抗癌药物。先进的微机械加工技术为高质量凹型微孔的制备提供了一种极好的技术解决方案。
A novel approach based on advanced micromachining is demonstrated to fabricate concave microwell arrays for the formation of high quality multicellular tumor spheroids. Microfabricated molds were prepared using a state-of-the-art CNC machining center, containing arrays of 3D convex micropillars with size ranging from 150 mu m to 600 mu m. Microscopic imaging of the micropillars machined on the mold showed smooth, curved microfeatures of a dramatic 3D shape. Agarose microwells could be easily replicated from the metallic molds. EMT-6 tumor cells seeded in the primary macrowell sedimented efficiently to the bottom of the concave microwells and formed multicellular spheroids within 48 h. Dense and homogeneous multicellular spheroids were obtained after 10 days of culture, confirming the suitability of the proposed approach. To facilitate long term spheroid culture and reliable on-chip drug assay, polydimethylsiloxane microwells were also replicated from the metallic molds. A solvent swelling method was adapted and optimized to Pluronic F127 towards physically entrapping the block copolymer molecules within the polydimethylsiloxane network and in turn to improve long term cell-binding resistance. Homogeneous multicellular spheroids were efficiently formed in the concave microwells and on-chip drug assays could be reliably carried out using curcumin as a model anti-cancer drug. Advanced micromachining provides an excellent technological solution to the fabrication of high quality concave microwells.