Publications

Fully Automated One-Step Production of Functional 3D Tumor Spheroids for High-Content Screening
François Monjaret / Mathieu Fernandes / Eve Duchemin-Pelletier / Amelie Argento / Sébastien Degot / and Joanne Young
21 September 2015
Keywords:
high-content screening, Micropatterns, 3D spheroids, assay development

Adoption of spheroids within high-content screening (HCS) has lagged behind high-throughput screening (HTS) due to issues with running complex assays on large three-dimensional (3D) structures.
To enable multiplexed imaging and analysis of spheroids, different cancer cell lines were grown in 3D on micropatterned 96-well plates with automated production of nine uniform spheroids per well. Spheroids achieve diameters of up to 600 µm, and reproducibility was experimentally validated (interwell and interplate CVdiameter <5%). Biphoton imaging confirmed that micropatterned spheroids exhibit characteristic cell heterogeneity with distinct microregions. Furthermore, central necrosis appears at a consistent spheroid size, suggesting standardized growth. Using three reference compounds (fluorouracil, irinotecan, and staurosporine), we validated HT-29 micropatterned spheroids on an HCS platform, benchmarking against hanging-drop spheroids. Spheroid formation and imaging in a single plate accelerate assay workflow, and fixed positioning prevents structures from overlapping or sticking to the well wall, augmenting image processing reliability. Furthermore, multiple spheroids per well increase the statistical confidence sufficiently to discriminate compound mechanisms of action and generate EC50 values for endpoints of cell death, architectural change, and size within a single-pass read. Higher quality data and a more efficient HCS work chain should encourage integration of micropatterned spheroid models within fundamental research and drug discovery applications.

Primary Bovine Extra-Embryonic Cultured Cells: A New Resource for the Study of In Vivo Peri-Implanting Phenotypes and Mesoderm Formation.
Hue I / Evain-Brion D / Fournier T / Degrelle SA
23 June 2015
Keywords:
Mesoderm, trophoblast, endoderm, bovine, gene expression, embryo

PLoS One. 2015 Jun 12;10(6):e0127330. doi: 10.1371/journal.pone.0127330. eCollection 2015.

 

Nuclear translocation of Cyclin B1 marks the restriction point for terminal cell cycle exit in G2 phase.
Müllers E / Silva Cascales H / Jaiswal H / Saurin AT / Lindqvist A.
23 June 2015
Keywords:
cell cycle, checkpoint recovery, Cyclin B1, DNA damage response, G2 phase, nuclear translocation recovery competence, senescence

Cell Cycle. 2014;13(17):2733-43. doi: 10.4161/15384101.2015.945831

 

Matrix stiffness drives epithelial–mesenchymal transition and tumour metastasis through a TWIST1–G3BP2 mechanotransduction pathway
Wei SC / Fattet L / Tsai JH / Guo Y / Pai VH / Majeski HE / Chen AC / Sah RL / Taylor SS / Engler AJ / Yang J
23 June 2015
Keywords:
EMT, cancer, mechanotransduction

Nat Cell Biol. 2015 May;17(5):678-88. doi: 10.1038/ncb3157. Epub 2015 Apr 20.

 

Cell-intrinsic adaptation of lipid composition to local crowding drives social behaviour.
Frechin M / Stoeger T / Daetwyler S / Gehin C / Battich N / Damm EM / Stergiou L / Riezman H / Pelkmans L
23 June 2015
Keywords:
Signaling, mechanotransduction, FAK, collective behavior, membrane composition

Nature. 2015 May 25. doi: 10.1038/nature14429.

 

Mechanical coupling between transsynaptic N-cadherin adhesions and actin flow stabilizes dendritic spines
Chazeau A / Garcia M / Czöndör K / Perrais D / Tessier B / Giannone 1 / Thoumine O
23 June 2015
Keywords:
Dendritic spines, synapse, N-cadherin, cytoskeleton, live cell imaging, filopodia

Mol Biol Cell. 2015 Mar 1;26(5):859-73. doi: 10.1091/mbc.E14-06-1086. Epub 2015 Jan 7.

 

Changes in actin dynamics and F-actin structure both in synaptoneurosomes of LRRK2(R1441G) mutant mice and in primary human fibroblasts of LRRK2(G2019S) mutation carriers
Caesar M / Felk S / Aasly JO / Gillardon F
23 June 2015
Keywords:
LRRK2; F-actin; synaptoneurosomes; fibroblasts; Parkinson’s disease; filopodia

Neuroscience. 2015 Jan 22;284:311-24. doi: 10.1016/j.neuroscience.2014.09.070. Epub 2014 Oct 6.

 

Micropattern size‐dependent endothelial differentiation from a human induced pluripotent stem cell line
Kusuma S / Smith Q / Facklam A / Gerecht S
23 June 2015
Keywords:
Differenciation, endothelial, stem cell, IPS, co-culture, differenciation ratio, perycytes, colony

J Tissue Eng Regen Med. 2015 Jan 30. doi: 10.1002/term.1985

 

Micropatterned Macrophage Analysis Reveals Global Cytoskeleton Constraints Induced by Bacillus anthracis Edema Toxin.
Trescos Y / Tessier E / Rougeaux C / Goossens PL / Tournier JN
23 June 2015
Keywords:
macrophage, infection, cytoskeleton, bacillus anthracis, BMDM, edema toxin

Infect Immun. 2015 May 26. pii: IAI.00479-15.

 

An in vitro one-dimensional assay to study growth factor-regulated tumor cell-macrophage interaction.
Sharma VP / Beaty BT / Cox D / Condeelis JS / Eddy RJ.
23 June 2015
Keywords:
cell motility, tumor, macrophage

Methods Mol Biol. 2014;1172:115-23. doi: 10.1007/978-1-4939-0928-5_10.

 

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