A cell-loss-free concave microwell array based size-controlled multi-cellular tumoroid generation for anti-cancer drug screening

Lee, Sang Woo; Jeong, Soo Yeon; Shin, Tae Hoon; Min, Junhong; Lee, Dong-Hyun; Jeong, Gi Seok

doi:10.1371/journal.pone.0219834

Cited by 19 publications

(15 citation statements)

References 47 publications

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“…For example, Liu et al used an ice lithography method carried out on superhydrophobic substrates to fabricate a quasi-spherical microwell chip 52 . Lee et al also made a concave microwell chip using a double PDMS casting procedure from the PMMA mold that was made by a laser carving machine 53 . In a creative and simple method, Kim et al fabricated a concave-bottomed microwell chip using the capillary action of liquid polymer on the pins of a computer CPU, which is achieved without expensive materials or complicated procedures 54 .…”

Section: Resultsmentioning

confidence: 99%

A combined 3D printing/CNC micro-milling method to fabricate a large-scale microfluidic device with the small size 3D architectures: an application for tumor spheroid production

Behroodi

Latifi

Bagheri

et al. 2020

Sci Rep

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The fabrication of a large-scale microfluidic mold with 3D microstructures for manufacturing of the conical microwell chip using a combined projection micro-stereolithography (PµSL) 3D printing/CNC micro-milling method for tumor spheroid formation is presented. The PµSL technique is known as the most promising method of manufacturing microfluidic chips due to the possibility of creating complex three-dimensional microstructures with high resolution in the range of several micrometers. The purpose of applying the proposed method is to investigate the influence of microwell depths on the formation of tumor spheroids. In the conventional methods, the construction of three-dimensional microstructures and multi-height chips is difficult, time-consuming, and is performed using a multi-step lithography process. Microwell depth is an essential parameter for microwell design since it directly affects the shear stress of the fluid flow and the diffusion of nutrients, respiratory gases, and growth factors. In this study, a chip was made with microwells of different depth varying from 100 to 500 µm. The mold of the microwell section is printed by the lab-made PµSL printer with 6 and 1 µm lateral and vertical resolutions. Other parts of the mold, such as the main chamber and micro-channels, were manufactured using the CNC micro-milling method. Finally, different parts of the master mold were assembled and used for PDMS casting. The proposed technique drastically simplifies the fabrication and rapid prototyping of large-scale microfluidic devices with high-resolution microstructures by combining 3D printing with the CNC micro-milling method.

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Section: Resultsmentioning

confidence: 99%

A combined 3D printing/CNC micro-milling method to fabricate a large-scale microfluidic device with the small size 3D architectures: an application for tumor spheroid production

Behroodi

Latifi

Bagheri

et al. 2020

Sci Rep

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show abstract

“…The tumoroids had dimensions that could provide tumor microenvironment. [13] In a study reported earlier the 80,000 μm 2 MCTS was obtained post 30 days of cell seeding, which the authors classified as the largest MCTS. [14] This size of tumoroids shows distinct zones of the heterogeneous population and the grape shaped 3D tumoroids recapitulates the nodal structure of solid tumors by mimicking various such aspects of nutrient gradient, [15] pH, [16] and Hypoxia.…”

Section: Tumoroid Formation Aided Through MDC Hydrogelmentioning

confidence: 98%

Differential Migration and Proliferation Potential of the Hydrogel Aided 3D Tumoroid

Modi

Kedaria

Vasita

2022

Macromolecular Bioscience

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For substantial in vitro cancer biology research, the 3D cell culture method has now been regarded as more suitable model expected to be recapitulating maximum in vivo tumor mass relevance. Despite of available techniques to develop in vitro 3D models, a system availing a physiologically relevant in vitro 3D model of primary lung adenocarcinoma with extracellular matrix (ECM) mimicry and similar tumorigenic properties still remains a quest. Thus, in the present study, chemically modified Dextran-Chitosan (MDC) hydrogel has been developed as a 3D tumoroid aiding scaffold. The 3D A549 tumoroids aided by the MDC scaffold have physiologically relevant proliferation, migration, invasive potential, and Gefitinib [targeting epidermal growth factor receptor (EGFR)] efficacy as compared to the 2D cultured cells. The surface topography and wettability of hydrogel availed in vivo micro tumor mass mimicking Lung adenocarcinoma 3D in vitro model. Thus, opening an innovative avenue for elucidating the disease mechanism and drug efficacy on relevant 3D cancer models in vitro.

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“…The fabrication process for the PDMS-based cell-lossfree (CLF) concave microwell array (1 mm microwell size, 225 wells/microwell array) was described in detail in our previous study [22]. The CLF concave microwell array was fabricated by using a 35 mm culture plate as the size slab.…”

Section: Cell Culture and Formation Of 3d Mcts-ecm Tumor Graftsmentioning

confidence: 99%

“…These MCT models are composed of nonmigrating cells, including an epithelial phenotype with apical-basal polarity by active cell-cell interactions based on the basement membrane. Our previous study also introduced the development of a CLF microwell array for the formation of robust MCTs using this array and suggested their usefulness through a drug response test [22]. However, to more accurately explain the therapeutic effect, it is necessary to consider cellular behavior, such as the motility of cancer cells through the ECM, which is a structural component constituting the in vivo TME, and the increase in drug resistance.…”

Section: Development Of a Macroscale 3d In Vitro Tumor Graft Model By...mentioning

confidence: 99%

Sporadic cell death in macroscale 3D tumor grafts with high drug resistance by activating cell-ECM interactions

2021

Self Cite

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In the tumor microenvironment (TME), the extracellular matrix (ECM) provides a dynamic structure for cell adhesion and cancer cell motility, such as migration and invasion, as well as remodeling. Matrix metalloproteinases (MMPs) promote cancer cell motility, which contributes to inducing drug resistance and thereby acquiring aggressive features. The drug resistance-induced 3D in vitro tumor model can be an effective model for therapeutic strategies for anticancer drugs targeting aggressive cancer cells. Here, we describe highly drug-resistant multicellular tumoroids (MCTs)-ECM tumor grafts under a macroscale dense 3D in vitro model through a combination of numerous MCTs and a collagen matrix. MCTs-ECM tumor grafts promote the high activity of MMP2 and MMP9 compared to general MCTs and induced cancer cell motility. Then, after the administration of anticancer drugs, the tumor grafts show increased drug resistance, with both the sporadic distribution of necrotic cells and the reduction of apoptotic portions, by activating cancer cell motility. MCTs-ECM tumor graft could be useful as a macroscale tumor graft model for inducing drug resistance by activating cancer cell motility and evaluating the efficacy of anticancer drugs targeting cancer with aggressive features.

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A cell-loss-free concave microwell array based size-controlled multi-cellular tumoroid generation for anti-cancer drug screening

Cited by 19 publications

References 47 publications

A combined 3D printing/CNC micro-milling method to fabricate a large-scale microfluidic device with the small size 3D architectures: an application for tumor spheroid production

A combined 3D printing/CNC micro-milling method to fabricate a large-scale microfluidic device with the small size 3D architectures: an application for tumor spheroid production

Differential Migration and Proliferation Potential of the Hydrogel Aided 3D Tumoroid

Sporadic cell death in macroscale 3D tumor grafts with high drug resistance by activating cell-ECM interactions

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