A 3D Printed Hanging Drop Dripper for Tumor Spheroids Analysis Without Recovery

Zhao, Liang; Xiu, Jidong; Liu, Yang; Zhang, Tianye; Pan, Wenjie; Zheng, Xiaonan; Zhang, Xueji

doi:10.1038/s41598-019-56241-0

Cited by 76 publications

(54 citation statements)

References 44 publications

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“…Moreover, the hanging drop method can originate spheroids with heterogeneous sizes and morphologies, impacting the spheroid standardization, which is essential for new drug screening. Recently, studies have developed different tools to minimize these limitations and facilitated the realization of this method [75][76][77]. For instance, the pressure-assisted network for droplet accumulation (PANDA) system consists of a pressure chip capable to create homogeneous and compact hanging drop array, enabling the fast and economical production of spheroids [75].…”

Section: Hanging Dropmentioning

confidence: 99%

Three-Dimensional Spheroids as In Vitro Preclinical Models for Cancer Research

et al. 2020

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Most cancer biologists still rely on conventional two-dimensional (2D) monolayer culture techniques to test in vitro anti-tumor drugs prior to in vivo testing. However, the vast majority of promising preclinical drugs have no or weak efficacy in real patients with tumors, thereby delaying the discovery of successful therapeutics. This is because 2D culture lacks cell–cell contacts and natural tumor microenvironment, important in tumor signaling and drug response, thereby resulting in a reduced malignant phenotype compared to the real tumor. In this sense, three-dimensional (3D) cultures of cancer cells that better recapitulate in vivo cell environments emerged as scientifically accurate and low cost cancer models for preclinical screening and testing of new drug candidates before moving to expensive and time-consuming animal models. Here, we provide a comprehensive overview of 3D tumor systems and highlight the strategies for spheroid construction and evaluation tools of targeted therapies, focusing on their applicability in cancer research. Examples of the applicability of 3D culture for the evaluation of the therapeutic efficacy of nanomedicines are discussed.

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Section: Hanging Dropmentioning

confidence: 99%

Three-Dimensional Spheroids as In Vitro Preclinical Models for Cancer Research

et al. 2020

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“…The physiological conditions such as continuous supply of nutrients, gas exchange, and vascularization are all important for closer recapitulation of in vivo environments. Recently, engineered microsystems integrated with optical tweezers, [ 240,241 ] acoustic waves, [ 242 ] electrodes, [ 243 ] and 3D printed architectures [ 244 ] have emerged as promising analytical platforms for providing deep insights on complex cellular interactions. Many of these microengineering systems, however, use PDMS as the base material, are often limited owing to its non‐degradable properties.…”

Section: Discussionmentioning

confidence: 99%

Engineered Microsystems for Spheroid and Organoid Studies

Kang

Kim

Lee

et al. 2020

Adv Healthcare Materials

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3D in vitro model systems such as spheroids and organoids provide an opportunity to extend the physiological understanding using recapitulated tissues that mimic physiological characteristics of in vivo microenvironments. Unlike 2D systems, 3D in vitro systems can bridge the gap between inadequate 2D cultures and the in vivo environments, providing novel insights on complex physiological mechanisms at various scales of organization, ranging from the cellular, tissue‐, to organ‐levels. To satisfy the ever‐increasing need for highly complex and sophisticated systems, many 3D in vitro models with advanced microengineering techniques have been developed to answer diverse physiological questions. This review summarizes recent advances in engineered microsystems for the development of 3D in vitro model systems. The relationship between the underlying physics behind the microengineering techniques, and their ability to recapitulate distinct 3D cellular structures and functions of diverse types of tissues and organs are highlighted and discussed in detail. A number of 3D in vitro models and their engineering principles are also introduced. Finally, current limitations are summarized, and perspectives for future directions in guiding the development of 3D in vitro model systems using microengineering techniques are provided.

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“…Sphericity remained stable around 90% and 75% for L929 and SKOV spheroids, respectively. The volume increase over time of spheroids correlates with an initial compaction phase followed by the proliferative phase of cancerous cells [37].…”

Section: Linear Correlation Between Sytox Intensity and Cell Densitymentioning

confidence: 99%

A 3D Cell Death Assay to Quantitatively Determine Ferroptosis in Spheroids

Demuynck

Efimova

Lin

et al. 2020

Cells

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The failure of drug efficacy in clinical trials remains a big issue in cancer research. This is largely due to the limitations of two-dimensional (2D) cell cultures, the most used tool in drug screening. Nowadays, three-dimensional (3D) cultures, including spheroids, are acknowledged to be a better model of the in vivo environment, but detailed cell death assays for 3D cultures (including those for ferroptosis) are scarce. In this work, we show that a new cell death analysis method, named 3D Cell Death Assay (3DELTA), can efficiently determine different cell death types including ferroptosis and quantitatively assess cell death in tumour spheroids. Our method uses Sytox dyes as a cell death marker and Triton X-100, which efficiently permeabilizes all cells in spheroids, was used to establish 100% cell death. After optimization of Sytox concentration, Triton X-100 concentration and timing, we showed that the 3DELTA method was able to detect signals from all cells without the need to disaggregate spheroids. Moreover, in this work we demonstrated that 2D experiments cannot be extrapolated to 3D cultures as 3D cultures are less sensitive to cell death induction. In conclusion, 3DELTA is a more cost-effective way to identify and measure cell death type in 3D cultures, including spheroids.

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A 3D Printed Hanging Drop Dripper for Tumor Spheroids Analysis Without Recovery

Cited by 76 publications

References 44 publications

Three-Dimensional Spheroids as In Vitro Preclinical Models for Cancer Research

Three-Dimensional Spheroids as In Vitro Preclinical Models for Cancer Research

Engineered Microsystems for Spheroid and Organoid Studies

A 3D Cell Death Assay to Quantitatively Determine Ferroptosis in Spheroids

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