Microfluidic channel-integrated hanging drop array chip operated by pushbuttons for spheroid culture and analysis

Park, Ju-Hwan; Kim, Hwisoo; Park, Je‐Kyun

doi:10.1039/d0an01091j

Cited by 14 publications

(9 citation statements)

References 31 publications

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“…To address this concern, open and bottomless hanging drop systems in microfluidics were developed with dynamic shear and interconnected networks for fluid flow; however, evaporation of liquid in hanging drops was still a limitation. The loss of culture media must be compensated by the frequent addition of liquid or equipped with other auxiliaries ( Park et al, 2020 ) to maintain the height of the suspended drop in the system. We addressed this challenge by utilizing a designed chamber or a spacer ( Supplementary Figure S1 ) to compensate for evaporative loss.…”

Section: Discussionmentioning

confidence: 99%

Flipped Well-Plate Hanging-Drop Technique for Growing Three-Dimensional Tumors

Jeong

Tin

Irudayaraj

2022

Front. Bioeng. Biotechnol.

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Three-dimensional (3D) tumor culture techniques are gaining popularity as in vitro models of tumoral tissue analogues. Despite the widespread interest, need, and present-day effort, most of the 3D tumor culturing methodologies have not gone beyond the inventors’ laboratories. This, in turn, limits their applicability and standardization. In this study, we introduce a straightforward and user-friendly approach based on standard 96-well plates with basic amenities for growing 3D tumors in a scaffold-free/scaffold-based format. Hanging drop preparation can be easily employed by flipping a universal 96-well plate. The droplets of the medium generated by the well-plate flip (WPF) method can be easily modified to address various mechanisms and processes in cell biology, including cancer. To demonstrate the applicability and practicality of the conceived approach, we utilized human colorectal carcinoma cells (HCT116) to first show the generation of large scaffold-free 3D tumor spheroids over 1.5 mm in diameter in single-well plates. As a proof-of-concept, we also demonstrate matrix-assisted tumor culture techniques in advancing the broader use of 3D culture systems. The conceptualized WPF approach can be adapted for a range of applications in both basic and applied biological/engineering research.

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Section: Discussionmentioning

confidence: 99%

Flipped Well-Plate Hanging-Drop Technique for Growing Three-Dimensional Tumors

Jeong

Tin

Irudayaraj

2022

Front. Bioeng. Biotechnol.

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“…Some use the microfluidic technology, which is useful owing to its simple operation and enables point-of-care testing. Through this method, Park et al, (2020) devised a finger-actuated microfluidic device for spheroid cultivation and analysis that facilitates programmed media exchange and media injection for further analysis [29]. Different sizes of BT474 spheroids were generated after seven days of growing and further analyzed in a LIVE/DEAD assay, indicating its spheroid growth in a manipulated ECM-mimicking environment.…”

Section: Hanging-drop Methodsmentioning

confidence: 99%

Recent Advances in Multicellular Tumor Spheroid Generation for Drug Screening

Lee

Kim

2021

Biosensors

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Multicellular tumor spheroids (MCTs) have been employed in biomedical fields owing to their advantage in designing a three-dimensional (3D) solid tumor model. For controlling multicellular cancer spheroids, mimicking the tumor extracellular matrix (ECM) microenvironment is important to understand cell–cell and cell–matrix interactions. In drug cytotoxicity assessments, MCTs provide better mimicry of conventional solid tumors that can precisely represent anticancer drug candidates’ effects. To generate incubate multicellular spheroids, researchers have developed several 3D multicellular spheroid culture technologies to establish a research background and a platform using tumor modelingvia advanced materials science, and biosensing techniques for drug-screening. In application, drug screening was performed in both invasive and non-invasive manners, according to their impact on the spheroids. Here, we review the trend of 3D spheroid culture technology and culture platforms, and their combination with various biosensing techniques for drug screening in the biomedical field.

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“…This prompts to generate models to study patients' immune response variability and drug sensitivity, and to characterize the interaction of immune cells with 3D CRC-derived spheroids [119,120,[125][126][127]136]. Tumor cell spheroids can be obtained applying different culture methods [125][126][127][128][129][130][131][132][133] (Figure 3). CRC is a very heterogeneous inter and intra patient disease [142][143][144][145].…”

Section: Crc Biological Features and Crc Spheroidsmentioning

confidence: 99%

“…Spheroids can be derived from either established cell lines or primary isolated tumor cells [ 35 , 118 , 119 , 120 , 121 , 122 , 123 , 124 , 125 , 126 , 127 , 128 ]. Usually, spheroids are obtained culturing tumor cells in ultra-low adherent plasticware or by hanging drop methods, forcing tumor cells to aggregate and generate tumor sphere in static or microfluidic systems [ 15 , 16 , 126 , 127 , 128 , 129 , 130 , 131 , 132 , 133 , 134 , 135 , 136 ]. The spheroids can be composed of a single (homotypic) or, less frequently, different (heterotypic) cell types [ 15 , 16 , 125 , 126 ].…”

Section: Tumor Cell Spheroids As a 3d Model To Study Tumor Cell Biology And Tumor-natural Killer Cross-talkmentioning

confidence: 99%

“…Usually, tumor cells are seeded onto ultra-low adherent culture wells [ 125 , 126 , 127 ]: this impairs adhesion of tumor cells to plastic substrate and favors the cell-to-cell aggregation. The size of spheroids can be controlled culturing the same number of cells in each well, and this is allowed when tumor cells are put in very little U or V bottomed wells or in hanging-drop plates [ 129 , 130 , 131 , 132 , 133 ]. Spheroids can be embedded into hydrogel drops that solidify at warm temperature, or by calcium addition, and be used as models of initial tumor growth [ 149 , 150 , 151 , 152 , 153 , 154 ].…”

Section: Tumor Cell Spheroids As a 3d Model To Study Tumor Cell Biology And Tumor-natural Killer Cross-talkmentioning

confidence: 99%

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

Poggi

Villa

Fernadez

et al. 2021

Cancers

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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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Microfluidic channel-integrated hanging drop array chip operated by pushbuttons for spheroid culture and analysis

Abstract: Although the hanging drop methods have a number of advantages for the spheroid culture, they suffer from reagent exchange procedures that depend on tedious and accurate liquid handling by manual...

Cited by 14 publications

References 31 publications

Flipped Well-Plate Hanging-Drop Technique for Growing Three-Dimensional Tumors

Flipped Well-Plate Hanging-Drop Technique for Growing Three-Dimensional Tumors

Recent Advances in Multicellular Tumor Spheroid Generation for Drug Screening

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

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