Assessing Cell Viability and Death in 3D Spheroid Cultures of Cancer Cells

Rolver, Michala G; Elingaard-Larsen, Line O; Pedersen, Stine Falsig

doi:10.3791/59714

Cited by 26 publications

(31 citation statements)

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“…The 3D cultures have specific properties and, therefore, their cultivation requires conditions that differ from 2D cultures ( Table 1 ) [ 54 ]. There are a number of methods for obtaining tumor spheroids.…”

Section: Methods For Obtaining Spheroids and Organoidsmentioning

confidence: 99%

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Promising Applications of Tumor Spheroids and Organoids for Personalized Medicine

Gilazieva

Ponomarev

Rutland

et al. 2020

Cancers

101

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One of the promising directions in personalized medicine is the use of three-dimensional (3D) tumor models such as spheroids and organoids. Spheroids and organoids are three-dimensional cultures of tumor cells that can be obtained from patient tissue and, using high-throughput personalized medicine methods, provide a suitable therapy for that patient. These 3D models can be obtained from most types of tumors, which provides opportunities for the creation of biobanks with appropriate patient materials that can be used to screen drugs and facilitate the development of therapeutic agents. It should be noted that the use of spheroids and organoids would expand the understanding of tumor biology and its microenvironment, help develop new in vitro platforms for drug testing and create new therapeutic strategies. In this review, we discuss 3D tumor spheroid and organoid models, their advantages and disadvantages, and evaluate their promising use in personalized medicine.

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Section: Methods For Obtaining Spheroids and Organoidsmentioning

confidence: 99%

“…Methocel prevents cell clumping and is inert whilst maintaining viscosity. This addition makes it possible to successfully generate more uniform spheroids and requires less cultivation time [ 54 , 55 ]. The hanging drop method is a simple, low cost, high performance method.…”

Section: Methods For Obtaining Spheroids and Organoidsmentioning

confidence: 99%

Promising Applications of Tumor Spheroids and Organoids for Personalized Medicine

Gilazieva

Ponomarev

Rutland

et al. 2020

Cancers

101

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“…3D cell culture was performed as described previously. 16 , 17 , 18 Briefly, 25 000 cells/well (embedded system) and 40 000 cells/well (on‐top system) were seeded into 24‐well plates. Cells were treated with C644‐0303 for 5 or 6 d, photographed every 2 d and quantified with ImageJ software.…”

Section: Methodsmentioning

confidence: 99%

C644‐0303, a small‐molecule inhibitor of the Wnt/β‐catenin pathway, suppresses colorectal cancer growth

Yan

Zhang

et al. 2021

Cancer Science

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The Wnt/β‐catenin signaling pathway plays an important role in tissue homeostasis, and its malignant activation is closely related to the occurrence and development of many cancers, especially colorectal cancer with adenomatous polyposis coli (APC) and CTNNB1 mutations. By applying a TCF/lymphoid‐enhancing factor (LEF) luciferase reporter system, the high‐throughput screening of 18 840 small‐molecule compounds was performed. A novel scaffold compound, C644‐0303, was identified as a Wnt/β‐catenin signaling inhibitor and exhibited antitumor efficacy. It inhibited both constitutive and ligand activated Wnt signals and its downstream gene expression. Functional studies showed that C644‐0303 causes cell cycle arrest, induces apoptosis, and inhibits cancer cell migration. Moreover, transcription factor array indicated that C644‐0303 could suppress various tumor‐promoting transcription factor activities in addition to Wnt/β‐catenin. Finally, C644‐0303 suppressed tumor spheroidization in a 3‐dimensional cell culture model and inhibited xenograft tumor growth in mice. In conclusion, we report a novel structural small molecular inhibitor targeting the Wnt/β‐catenin signaling pathway that has therapeutic potential for colorectal cancer treatment.

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“…3D cell culture models replicate the natural tumor architecture, thereby presenting an external proliferating zone, an internal quiescent zone with limited oxygen, nutrient and growth factor distribution, and a necrotic and hypoxic core [ 31 , 32 ]. All these factors might influence drug response.…”

Section: Pros and Cons Of The Current Models For Anti-cancer Drug Testingmentioning

confidence: 99%

3D Cell Culture Models as Recapitulators of the Tumor Microenvironment for the Screening of Anti-Cancer Drugs

Barbosa

Xavier

Pereira

et al. 2021

Cancers

119

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Today, innovative three-dimensional (3D) cell culture models have been proposed as viable and biomimetic alternatives for initial drug screening, allowing the improvement of the efficiency of drug development. These models are gaining popularity, given their ability to reproduce key aspects of the tumor microenvironment, concerning the 3D tumor architecture as well as the interactions of tumor cells with the extracellular matrix and surrounding non-tumor cells. The development of accurate 3D models may become beneficial to decrease the use of laboratory animals in scientific research, in accordance with the European Union’s regulation on the 3R rule (Replacement, Reduction, Refinement). This review focuses on the impact of 3D cell culture models on cancer research, discussing their advantages, limitations, and compatibility with high-throughput screenings and automated systems. An insight is also given on the adequacy of the available readouts for the interpretation of the data obtained from the 3D cell culture models. Importantly, we also emphasize the need for the incorporation of additional and complementary microenvironment elements on the design of 3D cell culture models, towards improved predictive value of drug efficacy.

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Assessing Cell Viability and Death in 3D Spheroid Cultures of Cancer Cells

Cited by 26 publications

References 0 publications

Promising Applications of Tumor Spheroids and Organoids for Personalized Medicine

Promising Applications of Tumor Spheroids and Organoids for Personalized Medicine

C644‐0303, a small‐molecule inhibitor of the Wnt/β‐catenin pathway, suppresses colorectal cancer growth

3D Cell Culture Models as Recapitulators of the Tumor Microenvironment for the Screening of Anti-Cancer Drugs

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