Leeds Institute of Medical Research at St. James's, St. James's University Hospital, University of Leeds, Beckett St., Leeds, West Yorkshire, LS9 7TF, UK.
Department of Mechanical Science and Engineering, University of Illinois at Urbana-Champaign, Urbana, IL, 61801, USA.
Sci Rep. 2020 Sep 28;10(1):15915. doi: 10.1038/s41598-020-72952-1.
Three-dimensional (3D) spheroidal cell cultures are now recognised as better models of cancers as compared to traditional cell cultures. However, established 3D cell culturing protocols and techniques are time-consuming, manually laborious and often expensive due to the excessive consumption of reagents. Microfluidics allows for traditional laboratory-based biological experiments to be scaled down into miniature custom fabricated devices, where cost-effective experiments can be performed through the manipulation and flow of small volumes of fluid. In this study, we characterise a 3D cell culturing microfluidic device fabricated from a 3D printed master. HT29 cells were seeded into the device and 3D spheroids were generated and cultured through the perfusion of cell media. Spheroids were treated with 5-Fluorouracil for five days through continuous perfusion and cell viability was analysed on-chip at different time points using fluorescence microscopy and Lactate dehydrogenase (LDH) assay on the supernatant. Increasing cell death was observed in the HT29 spheroids over the five-day period. The 3D cell culturing microfluidic device described in this study, permits on-chip anti-cancer treatment and viability analysis, and forms the basis of an effective platform for the high-throughput screening of anti-cancer drugs in 3D tumour spheroids.
三维(3D)球形细胞培养物现在被认为是比传统细胞培养更好的癌症模型。然而,由于试剂的大量消耗,现有的 3D 细胞培养方案和技术既耗时又费力,而且往往成本高昂。微流控技术允许将传统的基于实验室的生物学实验缩小到微型定制制造的设备中,通过操纵和流动小体积的流体,可以在这些设备中进行具有成本效益的实验。在这项研究中,我们对一种由 3D 打印模具制成的 3D 细胞培养微流控设备进行了表征。将 HT29 细胞接种到该设备中,并通过灌注细胞培养基来生成和培养 3D 球体。通过连续灌注将 5-氟尿嘧啶施用于球体 5 天,并使用荧光显微镜和上清液中的乳酸脱氢酶(LDH)测定法在不同时间点在芯片上分析细胞活力。在五天的时间内,观察到 HT29 球体中的细胞死亡增加。本研究中描述的 3D 细胞培养微流控设备允许在芯片上进行抗癌治疗和活力分析,并为在 3D 肿瘤球体中高通量筛选抗癌药物形成了有效的平台基础。