Three-Dimensional Aggregated Spheroid Model of Hepatocellular Carcinoma Using a 96-Pillar/Well Plate

Lee, Sang-Yun; Teng, Yun; Son, Miseol; Ku, Bosung; Hwang, Hyun Ju; Tergaonkar, Vinay; Chow, Pierce K. H.; Lee, Dong Woo; Nam, Do‐Hyun

doi:10.3390/molecules26164949

Cited by 16 publications

(11 citation statements)

References 27 publications

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“…Their spatial architecture promotes the establishment of diffusion gradients that could modify what it is seen in monolayer, where all cells are equally exposed to drugs ( 25 ). Our group has already published a work that supports the theory that OC cells tend to be more resistant to CDDP treatment when growing on 3D ( 17 ), and similar results have also been recently described for different drugs in 3D models of colorectal cancer ( 26 ), hepatocarcinoma ( 27 ), glioblastoma ( 28 ), breast cancer ( 29 ), melanoma ( 30 ), and also in OC ( 25 ).…”

Section: Discussionsupporting

confidence: 73%

Antitumoral Effect of Plocabulin in High Grade Serous Ovarian Carcinoma Cell Line Models

et al. 2022

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Ovarian cancer (OC) is a life-threatening tumor and the deadliest among gynecological cancers in developed countries. First line treatment with a carboplatin/paclitaxel regime is initially effective in the majority of patients, but most advanced OC will recur and develop drug resistance. Therefore, the identification of alternative therapies is needed. In this study, we employed a panel of high-grade serous ovarian cancer (HGSOC) cell lines, in monolayer and three-dimensional cell cultures. We evaluated the effects of a novel tubulin-binding agent, plocabulin, on proliferation, cell cycle, migration and invasion. We have also tested combinations of plocabulin with several drugs currently used in OC in clinical practice. Our results show a potent antitumor activity of plocabulin, inhibiting proliferation, disrupting microtubule network, and decreasing their migration and invasion capabilities. We did not observe any synergistic combination of plocabulin with cisplatin, doxorubicin, gemcitabine or trabectedin. In conclusion, plocabulin has a potent antitumoral effect in HGSOC cell lines that warrants further clinical investigation.

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Section: Discussionsupporting

confidence: 73%

Antitumoral Effect of Plocabulin in High Grade Serous Ovarian Carcinoma Cell Line Models

et al. 2022

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“…Successful cell migration was observed under three-dimensional cell culture conditions; analytical throughput was low due to the complex 3D cell culture process and difficulty in obtaining and analyzing images of cells that spread from the tumor area. A micropillar and microwell chip platform for 3D cell-based high throughput screening was previously developed by our group [9][10][11]. Measurement of the various 3D cell responses of many drugs is based on the 3D cell area exposed to drugs.…”

Section: Introductionmentioning

confidence: 99%

High Throughput 3D Cell Migration Assay Using Micropillar/Microwell Chips

Lee

Park²,

et al. 2022

Molecules

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The 3D cell migration assay was developed for the evaluation of drugs that inhibit cell migration using high throughput methods. Wound-healing assays have commonly been used for cell migration assays. However, these assays have limitations in mimicking the in vivo microenvironment of the tumor and measuring cell viability for evaluation of cell migration inhibition without cell toxicity. As an attempt to manage these limitations, cells were encapsulated with Matrigel on the surface of the pillar, and an analysis of the morphology of cells attached to the pillar through Matrigel was performed for the measurement of cell migration. The micropillar/microwell chips contained 532 pillars and wells, which measure the migration and viability of cells by analyzing the roundness and size of the cells, respectively. Cells seeded in Matrigel have a spherical form. Over time, cells migrate through the Matrigel and attach to the surface of the pillar. Cells that have migrated and adhered have a diffused shape that is different from the initial spherical shape. Based on our analysis of the roundness of the cells, we were able to distinguish between the diffuse and spherical shapes. Cells in Matrigel on the pillar that were treated with migration-inhibiting drugs did not move to the surface of the pillar and remained in spherical forms. During the conduct of experiments, 70 drugs were tested in single chips and migration-inhibiting drugs without cell toxicity were identified. Conventional migration assays were performed using transwell for verification of the four main migration-inhibiting drugs found on the chip.

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“…various 3D-HTS analysis data using pillar plates and micropillar/well chips [17][18][19][20][21][22] . In particular, an aggregated spheroid model (ASM) that maximized drug efficacy evaluation by recapitulating the tumor microenvironment was attempted and reported in hepatocellular carcinoma (HCC) 23 . Therefore, pillar plate-based 3D-ASM has the advantage of being able to mix ECM with cells as a scaffold-type 3D cell culture method, and stable 3D cell culture and research with high uniformity and reproducibility.…”

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confidence: 99%

Optimization of 3D-aggregated spheroid model (3D-ASM) for selecting high efficacy drugs

Lee

Hwang

Lee

2022

Sci Rep

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Various three-dimensional (3D) cell culture methods have been developed to implement tumor models similar to in vivo. However, the conventional 3D cell culture method has limitations such as difficulty in using an extracellular matrix (ECM), low experimental reproducibility, complex 3D cell culture protocol, and difficulty in applying to high array plates such as 96- or 384-plates. Therefore, detailed protocols related to robust 3D-aggregated spheroid model (3D-ASM) production were optimized and proposed. A specially designed wet chamber was used to implement 3D-ASM using the hepatocellular carcinoma (HCC) cell lines, and the conditions were established for the icing step to aggregate the cells in one place and optimized ECM gelation step. Immunofluorescence (IF) staining is mainly used to simultaneously analyze drug efficacy and changes in drug-target biomarkers. By applying the IF staining method to the 3D-ASM model, confocal microscopy imaging and 3D deconvolution image analysis were also successfully performed. Through a comparative study of drug response with conventional 2D-high throughput screening (HTS), the 3D-HTS showed a more comprehensive range of drug efficacy analyses for HCC cell lines and enabled selective drug efficacy analysis for the FDA-approved drug sorafenib. This suggests that increased drug resistance under 3D-HTS conditions does not reduce the analytical discrimination of drug efficacy, also drug efficacy can be analyzed more selectively compared to the conventional 2D-HTS assay. Therefore, the 3D-HTS-based drug efficacy analysis method using an automated 3D-cell spotter/scanner, 384-pillar plate/wet chamber, and the proposed 3D-ASM fabrication protocol is a very suitable platform for analyzing target drug efficacy in HCC cells.

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Three-Dimensional Aggregated Spheroid Model of Hepatocellular Carcinoma Using a 96-Pillar/Well Plate

Cited by 16 publications

References 27 publications

Antitumoral Effect of Plocabulin in High Grade Serous Ovarian Carcinoma Cell Line Models

Antitumoral Effect of Plocabulin in High Grade Serous Ovarian Carcinoma Cell Line Models

High Throughput 3D Cell Migration Assay Using Micropillar/Microwell Chips

Optimization of 3D-aggregated spheroid model (3D-ASM) for selecting high efficacy drugs

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