3D Cell Culture Systems: Tumor Application, Advantages, and Disadvantages

The traditional two-dimensional (2D) in vitro cell culture system (on a flat support) has long been used in cancer research. However, this system cannot be fully translated into clinical trials to ideally represent physiological conditions. This culture cannot mimic the natural tumor microenvironment due to the lack of cellular communication (cell-cell) and interaction (cell-cell and cell-matrix). To overcome these limitations, three-dimensional (3D) culture systems are increasingly developed in research and have become essential for tumor research, tissue engineering, and basic biology research. 3D culture has received much attention in the field of biomedicine due to its ability to mimic tissue structure and function. The 3D matrix presents a highly dynamic framework where its components are deposited, degraded, or modified to delineate functions and provide a platform where cells attach to perform their specific functions, including adhesion, proliferation, communication, and apoptosis. So far, various types of models belong to this culture: either the culture based on natural or synthetic adherent matrices used to design 3D scaffolds as biomaterials to form a 3D matrix or based on non-adherent and/or matrix-free matrices to form the spheroids. In this review, we first summarize a comparison between 2D and 3D cultures. Then, we focus on the different components of the natural extracellular matrix that can be used as supports in 3D culture. Then we detail different types of natural supports such as matrigel, hydrogels, hard supports, and different synthetic strategies of 3D matrices such as lyophilization, electrospiding, stereolithography, microfluid by citing the advantages and disadvantages of each of them. Finally, we summarize the different methods of generating normal and tumor spheroids, citing their respective advantages and disadvantages in order to obtain an ideal 3D model (matrix) that retains the following characteristics: better biocompatibility, good mechanical properties corresponding to the tumor tissue, degradability, controllable microstructure and chemical components like the tumor tissue, favorable nutrient exchange and easy separation of the cells from the matrix.

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A rhabdomyosarcoma hydrogel model to unveil cell-extracellular matrix interactions

Biomaterials Science ◽

10.1039/d1bm00929j ◽

2021 ◽

Author(s):

Mattia Saggioro ◽

Stefania D'Agostino ◽

Anna Gallo ◽

Sara Crotti ◽

Sara D'Aronco ◽

...

Keyword(s):

Cell Culture ◽

Extracellular Matrix ◽

Three Dimensional ◽

3D Culture ◽

3D Cell Culture ◽

Culture Systems ◽

Matrix Interactions ◽

In Vitro Systems ◽

Extracellular Matrix Interactions

Three-dimensional (3D) culture systems are progressively getting attention given their potential in overcoming limitations of the classical 2D in vitro systems. Among different supports for 3D cell culture, hydrogels (HGs)...

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Bioengineered 3D Glial Cell Culture Systems and Applications for Neurodegeneration and Neuroinflammation

SLAS DISCOVERY Advancing Life Sciences ◽

10.1177/2472555217691450 ◽

2017 ◽

Vol 22 (5) ◽

pp. 583-601 ◽

Cited By ~ 8

Author(s):

P. Marc D. Watson ◽

Edel Kavanagh ◽

Gary Allenby ◽

Matthew Vassey

Keyword(s):

Cell Culture ◽

Drug Discovery ◽

Glial Cell ◽

Critical Role ◽

3D Culture ◽

Cell Types ◽

Culture Systems ◽

Cellular Environment

Neurodegeneration and neuroinflammation are key features in a range of chronic central nervous system (CNS) diseases such as Alzheimer’s and Parkinson’s disease, as well as acute conditions like stroke and traumatic brain injury, for which there remains significant unmet clinical need. It is now well recognized that current cell culture methodologies are limited in their ability to recapitulate the cellular environment that is present in vivo, and there is a growing body of evidence to show that three-dimensional (3D) culture systems represent a more physiologically accurate model than traditional two-dimensional (2D) cultures. Given the complexity of the environment from which cells originate, and their various cell–cell and cell–matrix interactions, it is important to develop models that can be controlled and reproducible for drug discovery. 3D cell models have now been developed for almost all CNS cell types, including neurons, astrocytes, microglia, and oligodendrocyte cells. This review will highlight a number of current and emerging techniques for the culture of astrocytes and microglia, glial cell types with a critical role in neurodegenerative and neuroinflammatory conditions. We describe recent advances in glial cell culture using electrospun polymers and hydrogel macromolecules, and highlight how these novel culture environments influence astrocyte and microglial phenotypes in vitro, as compared to traditional 2D systems. These models will be explored to illuminate current trends in the techniques used to create 3D environments for application in research and drug discovery focused on astrocytes and microglial cells.

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Three-Dimensional Culture Models to Study Innate Anti-Tumor Immune Response: Advantages and Disadvantages

Cancers ◽

10.3390/cancers13143417 ◽

2021 ◽

Vol 13 (14) ◽

pp. 3417

Author(s):

Alessandro Poggi ◽

Federico Villa ◽

Jordi Leonardo Castrillo Fernadez ◽

Delfina Costa ◽

Maria Raffaella Zocchi ◽

...

Keyword(s):

Immune Response ◽

Innate Immunity ◽

Tumor Cell ◽

Nk Cells ◽

Tumor Cells ◽

Three Dimensional ◽

3D Culture ◽

Culture Systems ◽

Advantages And Disadvantages

Several approaches have shown that the immune response against tumors strongly affects patients’ clinical outcome. Thus, the study of anti-tumor immunity is critical to understand and potentiate the mechanisms underlying the elimination of tumor cells. Natural killer (NK) cells are members of innate immunity and represent powerful anti-tumor effectors, able to eliminate tumor cells without a previous sensitization. Thus, the study of their involvement in anti-tumor responses is critical for clinical translation. This analysis has been performed in vitro, co-incubating NK with tumor cells and quantifying the cytotoxic activity of NK cells. In vivo confirmation has been applied to overcome the limits of in vitro testing, however, the innate immunity of mice and humans is different, leading to discrepancies. Different activating receptors on NK cells and counter-ligands on tumor cells are involved in the antitumor response, and innate immunity is strictly dependent on the specific microenvironment where it takes place. Thus, three-dimensional (3D) culture systems, where NK and tumor cells can interact in a tissue-like architecture, have been created. For example, tumor cell spheroids and primary organoids derived from several tumor types, have been used so far to analyze innate immune response, replacing animal models. Herein, we briefly introduce NK cells and analyze and discuss in detail the properties of 3D tumor culture systems and their use for the study of tumor cell interactions with NK cells.

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Practical Applications of 3D Cell Culture Biotechnologies for Translational Oncology and Personalized Therapy

Biotekhnologiya ◽

10.21519/0234-2758-2020-36-3-3-15 ◽

2020 ◽

Vol 36 (3) ◽

pp. 3-15

Author(s):

D.A. Chudakova ◽

E.Yu. Skorova ◽

I.V. Reshetov

Keyword(s):

Cell Culture ◽

Extracellular Matrix ◽

3D Culture ◽

3D Cell Culture ◽

Personalized Therapy ◽

3D Bioprinting ◽

Culture Systems ◽

Culture Models ◽

2D And 3D

The creation of in vitro three-dimensional cellular model systems (in vitro 3D cultures) is a fast-growing leading-edge segment of the biotechnological industry. We have examined in this work the key 80 articles published after 2008, and focused on applications of in vitro 3D culture in translational oncology. We described a broad range of 3D culture systems, including models with and without extracellular matrix (ECM). 3D culture models based on decellularized ECM were discussed in more detail. The role of ECM in pathogeneis of malignant neoplasms, in particular, in the phenomenon of the tumor resistance to chemotherapy, was evaluated. 2D and 3D culture systems were compared, and natural and synthetic ECM were described, as well as the model creation based on 3D bioprinting. Particular attention was paid to in vitro models of various cancers, including those at the metastatic stage, based on 3D cell cultures, which maximally mimic the in vivo tumor behavior. The prospects of the practical application of 3D cell culture models in preclinical drag screening and in personalized therapy were discussed. We also presented our data on in vitro 2D and 3D culturing of human cells on various substrates. 3D cellular models, 3D bioprinting, biotechnology, extracellular matrix, cancer, translational medicine, personalized medicine, drag development, in vitro, ex vivo, oncology The authors are grateful to Dr. E. Shabalina for providing part of the experimental data and to OKA-Biotech Company for the samples of recombinant Funding-The work was supported by a Grant from the Russian Science Foundation (no. 18-15-00391). doi: 10.21519/0234-2758-2020-36-3-3-15

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Use of 3D culture systems to generate human induced pluripotent stem cell-derived [beta]-cells in vitro

Endocrine Abstracts ◽

10.1530/endoabs.57.003 ◽

2018 ◽

Author(s):

Fantuzzi Federica ◽

Toivonen Sanna ◽

Schiavo Andrea Alex ◽

Pachera Nathalie ◽

Rajaei Bahareh ◽

...

Keyword(s):

Stem Cell ◽

Beta Cells ◽

Pluripotent Stem Cell ◽

3D Culture ◽

Induced Pluripotent Stem Cell ◽

Culture Systems ◽

Induced Pluripotent

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Recent Progress in Prediction Systems for Drug-induced Liver Injury Using in vitro Cell Culture

Drug Metabolism Letters ◽

10.2174/1872312814666201202112610 ◽

2020 ◽

Vol 14 ◽

Author(s):

Shogo Ozawa ◽

Toshitaka Miura ◽

Jun Terashima ◽

Wataru Habano ◽

Seiichi Ishida

Keyword(s):

Cell Culture ◽

Recent Progress ◽

Inflammatory Cells ◽

Protein Adducts ◽

In Vitro Assays ◽

Drug Induced ◽

Drug Induced Liver Injury ◽

Culture Systems ◽

Cell Culture Systems

Background: In order to avoid drug-induced liver injury (DILI), in vitro assays, which enable the assessment of both metabolic activation and immune reaction processes that ultimately result in DILI, are needed. Objective: In this study, the recent progress in the application of in vitro assays using cell culture systems is reviewed for potential DILI-causing drugs/xenobiotics and a mechanistic study on DILI, as well as for the limitations of in vitro cell culture systems for DILI research. Methods: Information related to DILI was collected through a literature search of the PubMed database. Results: The initial biological event for the onset of DILI is the formation of cellular protein adducts after drugs have been metabolically activated by drug metabolizing enzymes. The damaged peptides derived from protein adducts lead to the activation of CD4+ helper T lymphocytes and recognition by CD8+ cytotoxic T lymphocytes, which destroy hepatocytes through immunological reactions. Because DILI is a major cause of drug attrition and drug withdrawal, numerous in vitro systems consisting of hepatocytes and immune/inflammatory cells, or spheroids of human primary hepatocytes containing non-parenchymal cells have been developed. These cellular-based systems have identified DILIinducing drugs with approximately 50% sensitivity and 90% specificity. Conclusion: Different co-culture systems consisting of human hepatocyte-derived cells and other immune/inflammatory cells have enabled the identification of DILI-causing drugs and of the actual mechanisms of action.

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Self-Assembling Polypeptide Hydrogels as a Platform to Recapitulate the Tumor Microenvironment

Cancers ◽

10.3390/cancers13133286 ◽

2021 ◽

Vol 13 (13) ◽

pp. 3286

Author(s):

Dariusz Lachowski ◽

Carlos Matellan ◽

Ernesto Cortes ◽

Alberto Saiani ◽

Aline F. Miller ◽

...

Keyword(s):

Cell Culture ◽

Tumor Microenvironment ◽

Cancer Cell ◽

Critical Role ◽

Hypoxia Inducible Factor ◽

Pancreatic Cancer Cell Line ◽

Cancer Cell Line ◽

Culture Systems ◽

A Cell

The tumor microenvironment plays a critical role in modulating cancer cell migration, metabolism, and malignancy, thus, highlighting the need to develop in vitro culture systems that can recapitulate its abnormal properties. While a variety of stiffness-tunable biomaterials, reviewed here, have been developed to mimic the rigidity of the tumor extracellular matrix, culture systems that can recapitulate the broader extracellular context of the tumor microenvironment (including pH and temperature) remain comparably unexplored, partially due to the difficulty in independently tuning these parameters. Here, we investigate a self-assembled polypeptide network hydrogel as a cell culture platform and demonstrate that the culture parameters, including the substrate stiffness, extracellular pH and temperature, can be independently controlled. We then use this biomaterial as a cell culture substrate to assess the effect of stiffness, pH and temperature on Suit2 cells, a pancreatic cancer cell line, and demonstrate that these microenvironmental factors can regulate two critical transcription factors in cancer: yes-associated protein 1 (YAP) and hypoxia inducible factor (HIF-1A).

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Mammary gland 3D cell culture systems in farm animals

Veterinary Research ◽

10.1186/s13567-021-00947-5 ◽

2021 ◽

Vol 52 (1) ◽

Author(s):

Laurence Finot ◽

Eric Chanat ◽

Frederic Dessauge

Keyword(s):

Cell Culture ◽

Mammary Gland ◽

Bacterial Infections ◽

Three Dimensional ◽

3D Cell Culture ◽

Farm Animals ◽

Culture Systems ◽

Cell Culture Systems

AbstractIn vivo study of tissue or organ biology in mammals is very complex and progress is slowed by poor accessibility of samples and ethical concerns. Fortunately, however, advances in stem cell identification and culture have made it possible to derive in vitro 3D “tissues” called organoids, these three-dimensional structures partly or fully mimicking the in vivo functioning of organs. The mammary gland produces milk, the source of nutrition for newborn mammals. Milk is synthesized and secreted by the differentiated polarized mammary epithelial cells of the gland. Reconstructing in vitro a mammary-like structure mimicking the functional tissue represents a major challenge in mammary gland biology, especially for farm animals for which specific agronomic questions arise. This would greatly facilitate the study of mammary gland development, milk secretion processes and pathological effects of viral or bacterial infections at the cellular level, all with the objective of improving milk production at the animal level. With this aim, various 3D cell culture models have been developed such as mammospheres and, more recently, efforts to develop organoids in vitro have been considerable. Researchers are now starting to draw inspiration from other fields, such as bioengineering, to generate organoids that would be more physiologically relevant. In this chapter, we will discuss 3D cell culture systems as organoids and their relevance for agronomic research.

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