Live cell division dynamics monitoring in 3D large spheroid tumor models using light sheet microscopy

Lorenzo, Corinne; Frongia, Céline; Jorand, Raphaël; Fehrenbach, Jérôme; Weiss, Pierre; Maandhui, Amina; Ducommun, Bernard; Lobjois, Valérie

doi:10.1186/1747-1028-6-22

Cited by 84 publications

(84 citation statements)

References 30 publications

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“…25 With these unique features, SPIM has been applied to image live cell proliferation processes in cellular spheroids. 23,26,27 However, most of microfluidics devices are not compatible with light sheet microscopy because of the geometries of the culture chambers or the arrangement of the flow channels.…”

Section: Introductionmentioning

confidence: 99%

Migration and vascular lumen formation of endothelial cells in cancer cell spheroids of various sizes

Patra

Peng²,

Peng

et al. 2014

Biomicrofluidics

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We developed a microfluidic device to culture cellular spheroids of controlled sizes and suitable for live cell imaging by selective plane illumination microscopy (SPIM). We cocultured human umbilical vein endothelial cells (HUVECs) within the spheroids formed by hepatocellular carcinoma cells, and studied the distributions of the HUVECs over time. We observed that the migration of HUVECs depended on the size of spheroids. In the spheroids of $200 lm diameters, HUVECs migrated outwards to the edges within 48 h; while in the spheroids of $250 lm diameters, there was no outward migration of the HUVECs up to 72 h. In addition, we studied the effects of pro-angiogenic factors, namely, vascular endothelial growth factor (VEGF) and fibroblast growth factor (b-FGF), on the migration of HUVECs in the carcinoma cell spheroid. The outward migration of HUVECs in 200 lm spheroids was hindered by the treatment with VEGF and b-FGF. Moreover, some of the HUVECs formed hollow lumen within 72 h under VEGF and b-FGF treatment. The combination of SPIM and microfluidic devices gives high resolution in both spatial and temporal domains. The observation of HUVECs in spheroids provides us insight on tumor vascularization, an ideal disease model for drug screening and fundamental studies.

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Section: Introductionmentioning

confidence: 99%

Migration and vascular lumen formation of endothelial cells in cancer cell spheroids of various sizes

Patra

Peng²,

Peng

et al. 2014

Biomicrofluidics

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“…Multi-photon imaging may be used to obtain a z-stack of a whole spheroid for 3D measurements 7 as this technology allows penetration of up to 500 μm, although some spheroids may still be too large to image whole spheroids via this method. Another option for imaging the whole spheroid is light-sheet-based microscopy, which allows visualization of the spheroid from multiple angles and penetrates up to 200 μm inside large spheroids 20,21 . Choice of imaging strategy may be influenced by the spheroid size, cell packing density and cell type.…”

Section: Discussionmentioning

confidence: 99%

Imaging- and Flow Cytometry-based Analysis of Cell Position and the Cell Cycle in 3D Melanoma Spheroids

Beaumont

Anfosso

Ahmed

et al. 2015

JoVE

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Three-dimensional (3D) tumor spheroids are utilized in cancer research as a more accurate model of the in vivo tumor microenvironment, compared to traditional two-dimensional (2D) cell culture. The spheroid model is able to mimic the effects of cell-cell interaction, hypoxia and nutrient deprivation, and drug penetration. One characteristic of this model is the development of a necrotic core, surrounded by a ring of G1 arrested cells, with proliferating cells on the outer layers of the spheroid. Of interest in the cancer field is how different regions of the spheroid respond to drug therapies as well as genetic or environmental manipulation. We describe here the use of the fluorescence ubiquitination cell cycle indicator (FUCCI) system along with cytometry and image analysis using commercial software to characterize the cell cycle status of cells with respect to their position inside melanoma spheroids. These methods may be used to track changes in cell cycle status, gene/protein expression or cell viability in different sub-regions of tumor spheroids over time and under different conditions. Video LinkThe video component of this article can be found at

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“…More recently, new imaging techniques like confocal and light sheet microscopy [100] have enabled 3-D imaging of tumor spheroids allowing researchers to monitor spheroidal uptake of fluorescent nanoparticles [101]. Despite these advances, scanning microscopy suffer from poor image resolution at higher penetration depth required for larger spheroids ( ) [100]. Magnetic resonance imaging (MRI) circumvents this issue by using magnetic radiation to image cellular proliferation noninvasively and has been utilized for not only imaging spheroidal morphology [102], [103] but also for monitoring intracellular pH [104].…”

Section: B Mcs Formation and Morphology Monitoringmentioning

confidence: 99%

“…Nevertheless, basic optical microscopy techniques are only capable of imaging the outer surfaces of spheroids without the need for freezing and sectioning, making real-time analysis impossible. More recently, new imaging techniques like confocal and light sheet microscopy [100] have enabled 3-D imaging of tumor spheroids allowing researchers to monitor spheroidal uptake of fluorescent nanoparticles [101]. Despite these advances, scanning microscopy suffer from poor image resolution at higher penetration depth required for larger spheroids ( ) [100].…”

Section: B Mcs Formation and Morphology Monitoringmentioning

confidence: 99%

From Cellular Cultures to Cellular Spheroids: Is Impedance Spectroscopy a Viable Tool for Monitoring Multicellular Spheroid (MCS) Drug Models?

Alexander

Price

Bhansali

2013

IEEE Rev. Biomed. Eng.

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The use of 3-D multicellular spheroid (MCS) models is increasingly being accepted as a viable means to study cell-cell, cell-matrix and cell-drug interactions. Behavioral differences between traditional monolayer (2-D) cell cultures and more recent 3-D MCS confirm that 3-D MCS more closely model the in vivo environment. However, analyzing the effect of pharmaceutical agents on both monolayer cultures and MCS is very time intensive. This paper reviews the use of electrical impedance spectroscopy (EIS), a label-free whole cell assay technique, as a tool for automated screening of cell drug interactions in MCS models for biologically/physiologically relevant events over long periods of time. EIS calculates the impedance of a sample by applying an AC current through a range of frequencies and measuring the resulting voltage. This review will introduce techniques used in impedance-based analysis of 2-D systems; highlight recently developed impedance-based techniques for analyzing 3-D cell cultures; and discuss applications of 3-D culture impedance monitoring systems.

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Live cell division dynamics monitoring in 3D large spheroid tumor models using light sheet microscopy

Cited by 84 publications

References 30 publications

Migration and vascular lumen formation of endothelial cells in cancer cell spheroids of various sizes

Migration and vascular lumen formation of endothelial cells in cancer cell spheroids of various sizes

Imaging- and Flow Cytometry-based Analysis of Cell Position and the Cell Cycle in 3D Melanoma Spheroids

From Cellular Cultures to Cellular Spheroids: Is Impedance Spectroscopy a Viable Tool for Monitoring Multicellular Spheroid (MCS) Drug Models?

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