A microfluidic platform for cultivating ovarian cancer spheroids and testing their responses to chemotherapies

Abstract

There is increasing interest in utilizing in vitro cultures as patient avatars to develop personalized treatment for cancer. Typical cultures utilize Matrigel-coated plates and media to promote the proliferation of cancer cells as spheroids or tumor explants. However, standard culture conditions operate in large volumes and require a high concentration of cancer cells to initiate this process. Other limitations include variability in the ability to successfully establish a stable line and inconsistency in the dimensions of these microcancers for in vivo drug response measurements. This paper explored the utility of microfluidics in the cultivation of cancer cell spheroids. Six patient-derived xenograft (PDX) tumors of high-grade serous ovarian cancer were used as the source material to demonstrate that viability and epithelial marker expression in the microfluidic cultures was superior to that of Matrigel or large volume 3D cultures. To further demonstrate the potential for miniaturization and multiplexing, we fabricated multichamber microfluidic devices with integrated microvalves to enable serial seeding of several chambers followed by parallel testing of several drug concentrations. These valve-enabled microfluidic devices permitted the formation of spheroids and testing of seven drug concentrations with as few as 100,000 cancer cells per device. Overall, we demonstrate the feasibility of maintaining difficul-to-culture primary cancer cells and testing drugs in a microfluidic device. This microfluidic platform may be ideal for drug testing and personalized therapy when tumor material is limited, such as following the acquisition of biopsy specimens obtained by fine-needle aspiration.

Using Spheroids as Building Blocks Towards 3D Bioprinting of Tumor Microenvironment

“…Other than alginate[ 98 ], hydrogels such as chitosan, thermosensitive gelatin, agarose, Matrigel, collagen, P(NIPAM-AA), photoinitiative gelatin methacrylate (gelMA), PEGDA, and hyaluronic acid-MA have all been examined for facilitating spheroid formation and growth[ 94 ]. In addition to droplet-based microfludics, lab-on-a-chip technology also can integrate hanging drop networks[ 103 - 105 ], microwell[ 50 , 106 ], U-shape microstructure, or micropillar into the platforms for spheroid formation and on-chip culture. Microfluidic platforms outperform conventional static culture methods through the introduction of a perfusion flow that could improve oxygen and nutrient transportation, sustaining long-term cell culture[ 107 , 108 ].…”

Modeling the Tumor Microenvironment of Ovarian Cancer: The Application of Self-Assembling Biomaterials

“…Overall, 15 drug combinations were identified to have potent cytotoxic properties. Dadgar et al demonstrated the potential of a serial and parallel perfused multi-chamber microfluidic device to test various drug concentrations on spheroids generated by low cellular input ( Figure 4 B) [ 104 ].…”

“…Spheroids were stained for the epithelial marker EpCAM, proliferation marker Ki-67 and nuclear marker DAPI ( bottom ). Scale bar: 50 µm (Reproduced with permission from [ 104 ], Copyright © (2020), Springer Nature). Poly-HEMA, poly 2-hydroxyethylmethacrylate; PDMS, polydimethylsiloxane.…”

Parallel and large‐scale antitumor investigation using stable chemical gradient and heterotypic three‐dimensional tumor coculture in a multi‐layered microfluidic device

“…These advances have provided several beneficial properties, such as facile and efficient operation, homogeneous and large-scale tumor production, and throughput monitoring. In addition, microfluidic tumor platforms have been widely applied for exploration of the tumor microenvironment, [16,24,30,31] tumor progression, [21,32] chemotherapy, [18][19][20]27,28,33] imaging, [25,34] and tumor-specific molecule detection. [35] However, microfluidic progress enabling the real-time detection and synchronous evaluation of 3D tumor responses to drugs at various concentrations by using a single device, was limited.…”