Institut Curie

7 articles published in JoVE

• Biochemistry

  
  Purification of Tubulin with Controlled Posttranslational Modifications and Isotypes from Limited Sources by Polymerization-Depolymerization Cycles Satish Bodakuntla*^1,2, A.S. Jijumon*^1,2, Carsten Janke^1,2, Maria M. Magiera^{1,2 1}PSL Research University, CNRS UMR3348, Institut Curie, ²Université Paris-Saclay, CNRS UMR3348, Université Paris Sud This protocol describes tubulin purification from small/medium-scale sources such as cultured cells or single mouse brains, using polymerization and depolymerization cycles. The purified tubulin is enriched in specific isotypes or has specific posttranslational modifications and can be used in in vitro reconstitution assays to study microtubule dynamics and interactions.

• Biology

  
  Quantifying Spatiotemporal Parameters of Cellular Exocytosis in Micropatterned Cells Hugo Lachuer^1,2,3, Pallavi Mathur^1,2,3, Kevin Bleakley⁴, Kristine Schauer^{1,2,3 1}Unité Mixte de Recherche 144 CNRS, Molecular Mechanisms of Intracellular Transport group, Institut Curie, 75005 Paris, France, ²PSL Research University, Paris, France, ³Sorbonne Université, Paris, France, ⁴INRIA, Université Paris-Sud, PSL Live imaging of lysosomal exocytosis on micropatterned cells allows a spatial quantification of this process. Morphology normalization using micropatterns is an outstanding tool to uncover general rules about the spatial distribution of cellular processes.

• Developmental Biology

  
  Stem cell-like Xenopus Embryonic Explants to Study Early Neural Developmental Features In Vitro and In Vivo Beatrice C. Durand^{1,2,3,4 1}Institut Curie, ²UMR 3387, CNRS, ³PSL Research University, ⁴Université Paris-Sud In Xenopus embryos, cells from the roof of the blastocoel are pluripotent and can be programmed to generate various tissues. Here, we describe protocols to use amphibian blastocoel roof explants as an assay system to investigate key in vivo and in vitro features of early neural development.

• Developmental Biology

  
  Protocols for Analyzing the Role of Paneth Cells in Regenerating the Murine Intestine using Conditional Cre-lox Mouse Models Lee Parry¹, Madeleine Young¹, Fatima El Marjou², Alan Richard Clarke^{1 1}European Cancer Stem Cell Research Institute, Cardiff University, ²Institut Curie Intestinal epithelial stem cells (ISCs) are intermingled with Paneth cells. These cells are differentiated progeny of the ISC, which support the ISCs and provide antibacterial protection. Here we demonstrate how we used transgenic conditional mouse models to establish that Paneth cells play a crucial role in maintaining the intestinal epithelia.

• Bioengineering

  
  Study of Cell Migration in Microfabricated Channels Pablo Vargas¹, Emmanuel Terriac², Ana-Maria Lennon-Duménil¹, Matthieu Piel^{2 1}Immunité et Cancer, Institut Curie, ²Compartimentation et Dynamique Cellulaires, Institut Curie A quantitative method to study spontaneous migration of cells in a one-dimensional confined microenvironment is described. This method takes advantage of microfabricated channels and can be used to study migration of large number of cells under different conditions in single experiments.

• Bioengineering

  
  Stretching Micropatterned Cells on a PDMS Membrane Nicolas Carpi¹, Matthieu Piel^{1 1}UMR 144, Institut Curie This manuscript presents a technique to apply or release forces on adherent cells or tissues using unidirectional stretching.

• Medicine

  
  Revealing the Cytoskeletal Organization of Invasive Cancer Cells in 3D Sara Geraldo¹, Anthony Simon¹, Danijela M. Vignjevic^{1 1}UMR 144 CNRS, Institut Curie This article presents a method to fluorescently label collagen that can be further used for both fix and live imaging of 3D cell cultures. We also provide an optimized protocol to visualize endogenous cytoskeletal proteins of cells cultured in 3D environments.