A multicellular spheroid-based sensor for anti-cancer therapeutics

Thielecke, Hagen; Mack, Alexandra; Robitzki, Andrea A.

doi:10.1016/s0956-5663(01)00140-3

Cited by 62 publications

(63 citation statements)

References 18 publications

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“…15,89,90 In both the microcavity and the microcapillary designs, it is important that the size of the cavity or capillary match with the size of spheroids, allowing the spheroids to be adequately analyzed by maintaining their shape and properties during measurements. 88 In these designs, Culture condition can be modified to include factors/proteins found in particular tumor microenvironment 17 Heterogeneous cell population are similar to cells of tumor that are in different phases of cell cycle including proliferating, hypoxic, and necrotic cells 24,53 The gene and protein expression levels of cells and thus the cellular behaviors are similar to in vivo levels 17,[27][28][29][30][31]40 Provide in vitro models for including different types of cells to build multicellular systems 52 Bridges the gap between in vitro and in vivo drug screening, possibly decreasing the use of animal models 5 …”

Section: D Cell Cultures In Cell-based Biosensorsmentioning

confidence: 99%

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Three-Dimensional Cell Culture Systems and Their Applications in Drug Discovery and Cell-Based Biosensors

Edmondson

Broglie

Adcock

et al. 2014

ASSAY and Drug Development Technologies

1,967

1,736

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Three-dimensional (3D) cell culture systems have gained increasing interest in drug discovery and tissue engineering due to their evident advantages in providing more physiologically relevant information and more predictive data for in vivo tests. In this review, we discuss the characteristics of 3D cell culture systems in comparison to the two-dimensional (2D) monolayer culture, focusing on cell growth conditions, cell proliferation, population, and gene and protein expression profiles. The innovations and development in 3D culture systems for drug discovery over the past 5 years are also reviewed in the article, emphasizing the cellular response to different classes of anticancer drugs, focusing particularly on similarities and differences between 3D and 2D models across the field. The progression and advancement in the application of 3D cell cultures in cell-based biosensors is another focal point of this review.

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Section: D Cell Cultures In Cell-based Biosensorsmentioning

confidence: 99%

“…89 Kloss et al demonstrated that drug-induced apoptosis in spheroids displayed increased impedance magnitudes, resulting from an altered morphology of the outer cells, using the microcavity system. 88 Hildebrandt et al used the capillary system to measure the impedance of mesenchymal stem cell (MSC) spheroids at three different conditions.…”

mentioning

confidence: 99%

Three-Dimensional Cell Culture Systems and Their Applications in Drug Discovery and Cell-Based Biosensors

Edmondson

Broglie

Adcock

et al. 2014

ASSAY and Drug Development Technologies

1,967

1,736

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“…[2][3][4] In the cosmetic field, such large-scale screens often utilize the enhanced green fluorescent protein (EGFP) gene as a reporter to allow sensitive, rapid, and convenient quantitation of gene expression.…”

Section: )mentioning

confidence: 99%

“…1) Recently, a biosensor was developed using 3D spheroid cultures and bioelectronically characterized by measuring physiological alterations after drug treatment. [2][3][4] In the cosmetic field, such large-scale screens often utilize the enhanced green fluorescent protein (EGFP) gene as a reporter to allow sensitive, rapid, and convenient quantitation of gene expression. 5) Although using 2D adherent cultures of melanin-producing cells is a useful method for the screening of skin-whitening ingredients, its utility is limited due to the time-consuming, laborious process, which only results in endpoint, static data with low sensitivity which requires multistep cell viability 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay for quantification.…”

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confidence: 99%

Development of Tyrosinase Promoter-Based Fluorescent Assay for Screening of Anti-melanogenic Agents

Lee

et al. 2015

Biological & Pharmaceutical Bulletin

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For screening of skin-whitening ingredients that modulate inhibition of melanogenesis, tyrosinase promoter-based assay using a three-dimensional (3D) spheroid culture technique is a beneficial tool to improve the accuracy of raw material screening in cosmetics through mimicking of the in vivo microenvironment. Although the advantages of high-throughput screening (HTS) are widely known, there has been little focus on specific cell-based promoter assays for HTS in identifying skin-whitening ingredients that inhibit accumulation of melanin. The aim of this study was therefore to develop a large-scale compatible assay through pTyr-EGFP, an enhanced green fluorescent protein (EGFP)-based tyrosinase-specific promoter, to seek potential melanogenesis inhibitors for cosmetic use. Herein, a stably transfected human melanoma cell line expressing EGFP under the control of a 2.2-kb fragment derived from the tyrosinase gene was generated. Spontaneous induction of the tyrosinase promoter by 3D spheroid culture resulted in increased expression of EGFP, providing a significant correlation with the tyrosinase mRNA level, and subsequent inhibition of tyrosinase activity. Importantly, the pTyr-EGFP system provided successful tracking of the changes in the live image and real-time monitoring. Thus tyrosinase promoter-based fluorescent assay using a 3D spheroid culture can be useful as a screening system for exploring the efficiency of anti-melanogenesis ingredients.Key words three-dimensional spheroid culture; tyrosinase promoter; fluorescent assay; high-throughput screening (HTS)Created with the purpose of mimicking in vivo environments, three-dimensional (3D) cultures have been shown to differ remarkably in the optimal cell growth, morphology, and differentiation compared with two-dimensional (2D) adherent cultures. Moreover, 3D cultures like spheroid cultures may come closer to the physiological interactions with neighboring cells and the extracellular matrix, providing valuable advanced tools for the evaluation of effectiveness. Particularly, therapeutic screening in 3D spheroid culture offers high potential for discovering drugs and biological relevances. 1)Recently, a biosensor was developed using 3D spheroid cultures and bioelectronically characterized by measuring physiological alterations after drug treatment. [2][3][4] In the cosmetic field, such large-scale screens often utilize the enhanced green fluorescent protein (EGFP) gene as a reporter to allow sensitive, rapid, and convenient quantitation of gene expression. 5)Although using 2D adherent cultures of melanin-producing cells is a useful method for the screening of skin-whitening ingredients, its utility is limited due to the time-consuming, laborious process, which only results in endpoint, static data with low sensitivity which requires multistep cell viability 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay for quantification. 6) We therefore developed a method to automatically monitor the activity of a tyrosinase promoter cloned upstream ...

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“…Two exceptions are the work by Poenick et al where a planar-type microcavity approach was used, with cells sitting on the microelectrodes, and that by Thielecket et al where spheroids were isolated in capillary chambers and then the impedance was measured using a commercial impedance analyzer; however, in both cases, the cultures were relatively large spheroids (∼300 µm diameter). 3,11,12 For many biological applications, conducting polymers have been shown to be preferable to traditional electronic materials due to properties such as chemical tunability, biocompatibility, and stability. 13 A very important property is their plasticity in terms of fabrication, a versatility which allows them to be used in a multitude of formats to fit different applications.…”

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confidence: 99%

Research Update: Electrical monitoring of cysts using organic electrochemical transistors

et al. 2015

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Organotypic three-dimensional (3D) cell culture models have the potential to act as surrogate tissues in vitro, both for basic research and for drug discovery/toxicology. 3D cultures maintain not only 3D architecture but also cell-cell and cell extracellular matrix interactions, particularly when grown in cysts or spheroids. Characterization of cell cultures grown in 3D formats, however, provides a significant challenge for cell biologists due to the incompatibility of these structures with commonly found optical or electronic monitoring systems. Electronic impedance spectroscopy is a cell culture monitoring technique with great potential; however, it has not been possible to integrate 3D cultures with commercially available systems to date. Cyst-like 3D cultures are particularly challenging due to their small size and difficulty in manipulation. Herein, we demonstrate isolation of cyst-like 3D cultures by capillarity and subsequent integration with the organic electrochemical transistor for monitoring the integrity of these structures. We show not only that this versatile device can be adapted to the cyst format for measuring resistance and, therefore, the quality of the cysts, but also can be used for quantitative monitoring of the effect of toxic compounds on cells in a 3D format. The ability to quantitatively predict effects of drugs on 3D cultures in vitro has large future potential for the fields of drug discovery and toxicology.

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A multicellular spheroid-based sensor for anti-cancer therapeutics

Cited by 62 publications

References 18 publications

Three-Dimensional Cell Culture Systems and Their Applications in Drug Discovery and Cell-Based Biosensors

Three-Dimensional Cell Culture Systems and Their Applications in Drug Discovery and Cell-Based Biosensors

Development of Tyrosinase Promoter-Based Fluorescent Assay for Screening of Anti-melanogenic Agents

Research Update: Electrical monitoring of cysts using organic electrochemical transistors

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