Generation and functional assessment of 3D multicellular spheroids in droplet based microfluidics platform.
Generation and functional assessment of 3D multicellular spheroids in droplet based microfluidics platform.
复制标题
基于液滴的微流体平台中 3D 多细胞球体的生成和功能评估。
DOI：
10.1039/c5lc01139f
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发表时间：
2016-02-07
期刊：
影响因子：
6.1
通讯作者：
Konry T
中科院分区：
文献类型：
--
作者：
Sabhachandani P;Motwani V;Cohen N;Sarkar S;Torchilin V;Konry T
在这里，我们描述了一种强大的微流体技术来生成和分析3D肿瘤球体，它类似于肿瘤微环境，可用作更有效的临床前药物测试和筛选模型。在聚二甲基硅氧烷（PDMS）微流控装置中产生单分散的载有细胞的藻酸盐液滴，所述装置结合联合收割机T形接头液滴产生和用于球体形成的外部凝胶化。所提出的方法有能力纳入多种细胞类型。为了我们研究的目的，我们用乳腺癌细胞系（MCF-7药物敏感和耐药）产生球状体，并将MCF-7的球状体与成纤维细胞系（HS-5）共培养。该设备能够在芯片上容纳1000个球状体，用于药物筛选和其他功能分析。通过将完全培养基连续灌注到装置中，将装置的阵列部分中的球状体的细胞活力维持两周。在我们的基于芯片的平台上分析了我们的3D肿瘤模型的功能性能以及标准化疗药物多柔比星（Dox）和标准药物组合Dox和Paclitaxel（PCT）的剂量依赖性反应。总之，我们的工作提供了一个简单而新颖的体外平台，用于生成、成像和分析均匀的3D单分散藻酸盐水凝胶肿瘤，用于各种Omic研究和治疗效率筛选，这是体内研究之前的重要转化步骤。
Here we describe a robust, microfluidic technique to generate and analyze 3D tumor spheroids, which resembles tumor microenvironment and can be used as a more effective preclinical drug testing and screening model. Monodisperse cell-laden alginate droplets were generated in Polydimethylsiloxane (PDMS) microfluidic devices that combine T-junction droplet generation and external gelation for spheroid formation. The proposed approach has the capability to incorporate multiple cell types. For the purposes of our study, we generated spheroids with breast cancer cell lines (MCF-7 drug sensitive and resistant) and co-culture spheroids of MCF-7 together with a fibroblast cell line (HS-5). The device has the capability to house 1000 spheroids on chip for drug screening and other functional analysis. Cellular viability of spheroids in the array part of the device was maintained for two weeks by continuous perfusion of complete media into the device. The functional performance of our 3D tumor models and a dose dependent response of standard chemotherapeutic drug, Doxorubicin (Dox) and standard drug combination Dox and Paclitaxel (PCT) was analyzed on our chip-based platform. Altogether, our work provides a simple and novel, in vitro platform to generate, image and analyze uniform, 3D monodisperse Alginate hydrogel tumors for various Omic studies and therapeutic efficiency screening, an important translational step before in vivo studies.