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\nFrancesca Antonelli  \nCitation: Antonelli, F. 3D Cell  \nModels in Radiobiology: Improving the Predictive Value of In Vitro Research. Int. J. Mol. Sci. 2023, 24, 10620. [https://doi.org/10.3390/](https://doi.org/10.3390/)[ ](https://doi.org/10.3390/)ijms241310620  \nAcademic Editor: Francis Cucinotta  \nReceived: 26 May 2023  \nRevised: 16 June 2023  \nAccepted: 21 June 2023  \nPublished: 25 June 2023  \nCopyright: © 2023 by the author. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license ([https://](https://)[ ](https://)[creativecommons.org/licenses/by/](creativecommons.org/licenses/by/)[ ](creativecommons.org/licenses/by/)[4.0/](4.0/)) .  \nLaboratory of Biomedical Technologies, Division of Health Protection Technologies, Agenzia Nazionale per le Nuove Tecnologie, l'Energia e lo Sviluppo Economico Sostenibile (ENEA), 00123 Rome, Italy; [francesca.antonelli@enea.it](francesca.antonelli@enea.it)  \nAbstract: Cancer is intrinsically complex, comprising both heterogeneous cellular composition and extracellular matrix. In vitro cancer research models have been widely used in the past to model and study cancer. Although two-dimensional (2D) cell culture models have traditionally been used for cancer research, they have many limitations, such as the disturbance of interactions between cellular and extracellular environments and changes in cell morphology, polarity, division mechanism, differentiation and cell motion. Moreover, 2D cell models are usually monotypic. This implies that 2D tumor models are ineffective at accurately recapitulating complex aspects of tumor cell growth, as well as their radiation responses. Over the past decade there has been significant uptake of threedimensional (3D) in vitro models by cancer researchers, highlighting a complementary model for studies of radiation effects on tumors, especially in conjunction with chemotherapy. The introduction of 3D cell culture approaches aims to model in vivo tissue interactions with radiation by positioning itself halfway between 2D cell and animal models, and thus opening up new possibilities in the study of radiation response mechanisms of healthy and tumor tissues.  \nKeywords: 3D cell models; Linear-quadratic model; dose–response curves; 3Dbioprinting; organoids; organ-on-a-chip; radiobiology  \n1. Introduction  \nTwo-dimensional (2D) cell cultures have historically been used for radiobiological studies and for modelling interactions between radiation and tissues, and so many of the dogmas of radiobiology are based on cellular and molecular responses of cells grown attached to a plastic surface [1–3] . In particular, the “gold standards” used to assess sensitivity towards radiotherapy are represented by the “clonogenic assay”, a test used to measure reproductive cell survival in vitro [4,5], and DNA damage evaluation in established cell lines [6,7] . Although these approaches are well-accepted, contributing significantly to the understanding of the mechanisms underlying cellular response to radiation, 2D systems depict an oversimplified representation of reality. For example, 2D cell cultures are usually monotypic, being made up of only a single cell type. In these models, the paracrine signaling between cells of different types is completely abolished. The tumor microenvironment, made up of both malignant cells and nonmalignant cellular and non-cellular components, can heavily condition the disease initiation, progression and treatment response of the tumor, since malignant cells and stromal components reciprocally communicate. Cancerassociated fibroblasts, as well as other tumor-associated cells, including endothelial cells and adipocytes [8], can highly increase tumor radioresistance through pro-survival factor secretion, immunomodulatory signals and contact-mediated s","cbCaipxK4gr3hjh9","https://ap.wps.com/l/cbCaipxK4gr3hjh9","pdf",2170420,25,"English","# Introduction\n## Limitations of 2D radiobiology models\n## Advancements in 3D cell culture technologies\n## Benefits and modeling of radiation responses","[{\"question\":\"Why are 2D cell culture models limited for radiobiology research?\",\"answer\":\"2D cultures simplify biology by disrupting interactions between cells and the extracellular environment, altering morphology and behaviors, and often lacking multicellular composition that supports accurate tumor microenvironment signaling.\"},{\"question\":\"How do 3D in vitro models improve predictive value compared with 2D models?\",\"answer\":\"3D approaches better position experiments between 2D systems and animal models, enabling more realistic modeling of tissue interactions with radiation and supporting more relevant tumor and healthy tissue response mechanisms.\"},{\"question\":\"What roles does the tumor microenvironment play in radiation response modeling?\",\"answer\":\"The microenvironment, including malignant and nonmalignant components such as cancer-associated fibroblasts and other stromal cells, can condition disease initiation, progression, and treatment response through signaling that can increase radioresistance.\"}]","3D Cell Models in Radiobiology - Improving the Predictive Value of In Vitro Research | PDF",1790562251,63]