Comparative Analysis of Dasatinib Effect between 2D and 3D Tumor Cell Cultures

Sabetta, Samantha; Vecchiotti, Davide; Clementi, Letizia; Nolfi, Mauro Di Vito; Zazzeroni, Francesca; Angelucci, Adriano

doi:10.3390/pharmaceutics15020372

Cited by 9 publications

(7 citation statements)

References 24 publications

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“…This phenomenon was evident in constructs consisting solely of tumor cells (3D-A431) and in tricellular tumor models (3D-cSCC). Over time, cellular aggregates appeared and merged into larger clusters, in line with previous research findings [6,[23][24][25]. The transition from smaller to larger clusters, encompassing the tumor core, was particularly noticeable in tricellular 3D-cSCC constructs.…”

Section: Discussionsupporting

confidence: 89%

3D modeling of normal skin and cutaneous squamous cell carcinoma. A comparative study in 2D cultures, spheroids, and 3D bioprinted systems

Kurzyk,

Szumera-Ciećkiewicz,

Miłoszewska

et al. 2024

Biofabrication

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The current cancer research and drug testing are primarily based on 2D cell cultures and animal models. However, these methods have limitations and yield distinct drug response patterns. This study addressed this gap by developing an innovative in vitro human 3D normal skin model and a multicellular model of human cutaneous squamous cell carcinoma (cSCC) using 3D bioprinting technology. Comparative analyzes were performed between bioprinted 3D-cSCC model, consisting of HaCaT keratinocytes, primary normal human dermal fibroblasts and A431 cancer cells (tricellular), bioprinted 3D-A431 model composed of A431 cancer cells only (monocellular), A431 cancer cell spheroids, and conventional 2D models. The models were structurally characterized by light microscopy, immunofluorescence (LIVE/DEAD assay, confocal microscopy) and immunohistochemistry (hematoxylin/eosin, p63, vimentin, Ki67, EGFR stainings). The spatial arrangement of the 3D models was analyzed using the ARIVIS Scientific Image Analysis Platform. All models were also functionally assessed by cetuximab (CTX) response testing with the MTS assay. 3D-cSCC models were maintained for up to 16 weeks. Morphological and histological examinations confirmed the presence of skin-like layers in the bioprinted 3D models of normal skin, and the intricate and diverse features of the bioprinted skin cancer model, replicating the critical in vivo characteristics. In both mono- and tricellular bioprinted tumor constructs, there was a gradual formation and continuous growth of spheroid-like clusters of cancer cells, significantly influencing the morphology of the models. Cancer cells in the 3D bioprinted constructs showed reduced sensitivity to CTX compared to spheroids and 2D cultures. This study underscores the potential of 3D multicellular models in elucidating drug responses and gaining a better understanding the intricate interplay of cellular components within the tumor microenvironment. Developing the multicellular 3D tumor model paves the way for new research critical to advancing fundamental cancer research and future clinical applications, particularly drug response testing.

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Section: Discussionsupporting

confidence: 89%

3D modeling of normal skin and cutaneous squamous cell carcinoma. A comparative study in 2D cultures, spheroids, and 3D bioprinted systems

Kurzyk,

Szumera-Ciećkiewicz,

Miłoszewska

et al. 2024

Biofabrication

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“…XTT assay results corroborated the live/dead staining findings, with an IC 50 value of 31.7 ± 1.4 nM for the xeno-free 3D model on day 5. Higher resistance against cytotoxicity of 3D bioprinted models compared to 2D cultures has been reported by a number of groups, including ours, for various models and drugs [72][73][74][75]. This finding is most likely due to the cells interacting in 3D networks compared to monolayer cultures, in which all cells have direct contact with the medium with their whole surface all the time.…”

Section: Toxicity Testingsupporting

confidence: 65%

Xeno-Free 3D Bioprinted Liver Model for Hepatotoxicity Assessment

Ali,

Berg,

Roehrs

et al. 2024

IJMS

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Three-dimensional (3D) bioprinting is one of the most promising methodologies that are currently in development for the replacement of animal experiments. Bioprinting and most alternative technologies rely on animal-derived materials, which compromises the intent of animal welfare and results in the generation of chimeric systems of limited value. The current study therefore presents the first bioprinted liver model that is entirely void of animal-derived constituents. Initially, HuH-7 cells underwent adaptation to a chemically defined medium (CDM). The adapted cells exhibited high survival rates (85–92%) after cryopreservation in chemically defined freezing media, comparable to those preserved in standard medium (86–92%). Xeno-free bioink for 3D bioprinting yielded liver models with high relative cell viability (97–101%), akin to a Matrigel-based liver model (83–102%) after 15 days of culture. The established xeno-free model was used for toxicity testing of a marine biotoxin, okadaic acid (OA). In 2D culture, OA toxicity was virtually identical for cells cultured under standard conditions and in CDM. In the xeno-free bioprinted liver model, 3-fold higher concentrations of OA than in the respective monolayer culture were needed to induce cytotoxicity. In conclusion, this study describes for the first time the development of a xeno-free 3D bioprinted liver model and its applicability for research purposes.

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“…Similarly, various head and neck squamous cell carcinoma cells were reported to show decreased sensitivity to cisplatin and cetuximab in 3D [ 51 ]. Prostate cancer cells DU145 and glioblastoma cells U87 were considerably more resistant to dasatinib-induced toxicity than corresponding cells cultured in monolayer, which was explained by the very limited ability of the drug to penetrate into the 3D model [ 52 ]. Thus, cells in 3D constructs are often observed to be less sensitive to drug treatment than monolayer cultures.…”

Section: Discussionmentioning

confidence: 99%

Generation of a Perfusable 3D Lung Cancer Model by Digital Light Processing

Mei

Berg

et al. 2023

IJMS

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Lung cancer still has one of the highest morbidity and mortality rates among all types of cancer. Its incidence continues to increase, especially in developing countries. Although the medical field has witnessed the development of targeted therapies, new treatment options need to be developed urgently. For the discovery of new drugs, human cancer models are required to study drug efficiency in a relevant setting. Here, we report the generation of a non-small cell lung cancer model with a perfusion system. The bioprinted model was produced by digital light processing (DLP). This technique has the advantage of including simulated human blood vessels, and its simple assembly and maintenance allow for easy testing of drug candidates. In a proof-of-concept study, we applied gemcitabine and determined the IC50 values in the 3D models and 2D monolayer cultures and compared the response of the model under static and dynamic cultivation by perfusion. As the drug must penetrate the hydrogel to reach the cells, the IC50 value was three orders of magnitude higher for bioprinted constructs than for 2D cell cultures. Compared to static cultivation, the viability of cells in the bioprinted 3D model was significantly increased by approximately 60% in the perfusion system. Dynamic cultivation also enhanced the cytotoxicity of the tested drug, and the drug-mediated apoptosis was increased with a fourfold higher fraction of cells with a signal for the apoptosis marker caspase-3 and a sixfold higher fraction of cells positive for PARP-1. Altogether, this easily reproducible cancer model can be used for initial testing of the cytotoxicity of new anticancer substances. For subsequent in-depth characterization of candidate drugs, further improvements will be necessary, such as the generation of a multi-cell type lung cancer model and the lining of vascular structures with endothelial cells.

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Comparative Analysis of Dasatinib Effect between 2D and 3D Tumor Cell Cultures

Cited by 9 publications

References 24 publications

3D modeling of normal skin and cutaneous squamous cell carcinoma. A comparative study in 2D cultures, spheroids, and 3D bioprinted systems

3D modeling of normal skin and cutaneous squamous cell carcinoma. A comparative study in 2D cultures, spheroids, and 3D bioprinted systems

Xeno-Free 3D Bioprinted Liver Model for Hepatotoxicity Assessment

Generation of a Perfusable 3D Lung Cancer Model by Digital Light Processing

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