High-content analysis of biomarker intensity and distribution in 3D microtissues

Waschow, Marcel; Letzsch, Stefan; Boettcher, Karin; Kelm, Jens M.

doi:10.1038/nmeth.f.359

Cited by 13 publications

(10 citation statements)

References 7 publications

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“…Given the intense interest recently shown in the area of 3D tumor models, the problem of the morphological heterogeneity of spherical colonies has already been addressed. Several authors advise monitoring different morphological parameters such as diameter, perimeter, area, volume and sphericity, the variability of which may affect the reproducibility of the results obtained 31 36 37 38 39 . However, a lack of quantitative analytical methods makes these observations purely theoretical.…”

Section: Discussionmentioning

confidence: 99%

3D tumor spheroid models for in vitro therapeutic screening: a systematic approach to enhance the biological relevance of data obtained

Zanoni

Piccinini

Arienti

et al. 2016

Sci Rep

842

710

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The potential of a spheroid tumor model composed of cells in different proliferative and metabolic states for the development of new anticancer strategies has been amply demonstrated. However, there is little or no information in the literature on the problems of reproducibility of data originating from experiments using 3D models. Our analyses, carried out using a novel open source software capable of performing an automatic image analysis of 3D tumor colonies, showed that a number of morphology parameters affect the response of large spheroids to treatment. In particular, we found that both spheroid volume and shape may be a source of variability. We also compared some commercially available viability assays specifically designed for 3D models. In conclusion, our data indicate the need for a pre-selection of tumor spheroids of homogeneous volume and shape to reduce data variability to a minimum before use in a cytotoxicity test. In addition, we identified and validated a cytotoxicity test capable of providing meaningful data on the damage induced in large tumor spheroids of up to diameter in 650 μm by different kinds of treatments.

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Section: Discussionmentioning

confidence: 99%

3D tumor spheroid models for in vitro therapeutic screening: a systematic approach to enhance the biological relevance of data obtained

Zanoni

Piccinini

Arienti

et al. 2016

Sci Rep

842

710

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“…Autoradiography reveals zones of heavily 3 H-FMISO labeled (white silver grains) live proliferating cells, intermediately labeled quiescent cells, and unlabeled (dark) necrotic core (Rasey et al , 1985). Perkin Elmer’s in vivo near-infrared (NIR) agents allow visualization of hypoxic areas and quantification of cancer-associated biomarkers in live tumor microtissue or spheroids (Waschow et al , 2012). Fluorescent-based probes allow detection of hypoxic regions within MCTS by confocal or fluorescent microscopy.…”

Section: Applications Of Tumor Spheroids In Cancer Researchmentioning

confidence: 99%

Three-dimensional culture systems in cancer research: Focus on tumor spheroid model

Nath

Devi

2016

Pharmacology & Therapeutics

702

667

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Cancer cells propagated in three-dimensional (3D) culture systems exhibit physiologically relevant cell-cell and cell-matrix interactions, gene expression and signaling pathway profiles, heterogeneity and structural complexity that reflect in vivo tumors. In recent years, development of various 3D models have improved the study of host-tumor interaction and use of high-throughput screening platforms for anti-cancer drug discovery and development. This review attempts to summarize the various 3D culture systems, with an emphasis on the most well characterized and widely applied model - multicellular tumor spheroids. This review also highlights the various techniques to generate tumor spheroids, methods to characterize them, and its applicability in cancer research.

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“…For carbonic anhydrase IX, at least in cancer tissues, probably the most strongly induced HIF target gene, a non-antibody-mediated fluorescent in vivo probe (called HypoxiSense 680) has been developed. 20 , 21 However, at best, these techniques provide only indirect evidence for tissue hypoxia due to self-adaptation, 22 “normoxic” regulation, and cell type-specific expression. 23 At least the latter point has been circumvented by the generation of transgenic mice ubiquitously and constitutively expressing a luciferase reporter gene fused to the O 2 -dependent degradation domain of HIF-1α.…”

Section: How Can the Tissue Po 2 Be Visualized?mentioning

confidence: 99%

Frequently asked questions in hypoxia research

Wenger¹,

Kurtcuoglu²,

Scholz³

et al. 2015

183

159

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“What is the O2 concentration in a normoxic cell culture incubator?” This and other frequently asked questions in hypoxia research will be answered in this review. Our intention is to give a simple introduction to the physics of gases that would be helpful for newcomers to the field of hypoxia research. We will provide background knowledge about questions often asked, but without straightforward answers. What is O2 concentration, and what is O2 partial pressure? What is normoxia, and what is hypoxia? How much O2 is experienced by a cell residing in a culture dish in vitro vs in a tissue in vivo? By the way, the O2 concentration in a normoxic incubator is 18.6%, rather than 20.9% or 20%, as commonly stated in research publications. And this is strictly only valid for incubators at sea level.

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High-content analysis of biomarker intensity and distribution in 3D microtissues

Cited by 13 publications

References 7 publications

3D tumor spheroid models for in vitro therapeutic screening: a systematic approach to enhance the biological relevance of data obtained

3D tumor spheroid models for in vitro therapeutic screening: a systematic approach to enhance the biological relevance of data obtained

Three-dimensional culture systems in cancer research: Focus on tumor spheroid model

Frequently asked questions in hypoxia research

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