Hisey Colin L, Mitxelena-Iribarren Oihane, Martínez-Calderón Miguel, Gordon Jaymeson B, Olaizola Santiago M, Benavente-Babace Ainara, Mujika Maite, Arana Sergio, Hansford Derek J
Department of Biomedical Engineering, The Ohio State University, Columbus, Ohio 43210, USA.
Department of Physics, University of Ottawa, Ottawa, Ontario K1N 6N5, Canada.
Biomicrofluidics. 2019 Jul 23;13(4):044105. doi: 10.1063/1.5103269. eCollection 2019 Jul.
Highly migratory cancer cells often lead to metastasis and recurrence and are responsible for the high mortality rates in many cancers despite aggressive treatment. Recently, the migratory behavior of patient-derived glioblastoma multiforme cells on microtracks has shown potential in predicting the likelihood of recurrence, while at the same time, antimetastasis drugs have been developed which require simple yet relevant high-throughput screening systems. However, robust platforms which can reliably seed single cells and measure their migration while mimicking the physiological tumor microenvironment have not been demonstrated. In this study, we demonstrate a microfluidic device which hydrodynamically seeds single cancer cells onto stamped or femtosecond laser ablated polystyrene microtracks, promoting 1D migratory behavior due to the cells' tendency to follow topographical cues. Using time-lapse microscopy, we found that single U87 glioblastoma multiforme cells migrated more slowly on laser ablated microtracks compared to stamped microtracks of equal width and spacing (p < 0.05) and exhibited greater directional persistence on both 1D patterns compared to flat polystyrene (p < 0.05). Single-cell morphologies also differed significantly between flat and 1D patterns, with cells on 1D substrates exhibiting higher aspect ratios and less circularity (p < 0.05). This microfluidic platform could lead to automated quantification of single-cell migratory behavior due to the high predictability of hydrodynamic seeding and guided 1D migration, an important step to realizing the potential of microfluidic migration assays for drug screening and individualized medicine.
高迁移性癌细胞常常导致转移和复发,并且尽管进行了积极治疗,仍是许多癌症高死亡率的原因。最近,患者来源的多形性胶质母细胞瘤细胞在微轨道上的迁移行为已显示出预测复发可能性的潜力,与此同时,已开发出抗转移药物,这需要简单而相关的高通量筛选系统。然而,尚未证明有能够可靠地接种单细胞并在模拟生理肿瘤微环境的同时测量其迁移的强大平台。在本研究中,我们展示了一种微流控装置,该装置通过流体动力学将单个癌细胞接种到压印或飞秒激光烧蚀的聚苯乙烯微轨道上,由于细胞倾向于遵循地形线索而促进一维迁移行为。使用延时显微镜,我们发现与宽度和间距相等的压印微轨道相比,单个U87多形性胶质母细胞瘤细胞在激光烧蚀微轨道上迁移得更慢(p < 0.05),并且与平坦聚苯乙烯相比,在两种一维模式上均表现出更大的方向持续性(p < 0.05)。平坦和一维模式之间的单细胞形态也存在显著差异,一维基质上的细胞表现出更高的纵横比和更低的圆形度(p < 0.05)。由于流体动力学接种的高可预测性和引导的一维迁移,这种微流控平台可导致单细胞迁移行为的自动量化,这是实现微流控迁移分析用于药物筛选和个性化医学潜力的重要一步。