Cambier, T., Honegger, T., Vanneaux, V., Berthier, J., Peyrade, D., Blanchoin, L., … Théry, M.
Laboratoire de Physiologie Cellulaire et Végétale, Institut de Recherche en Technologie et Science pour le Vivant, UMR5168, CEA, INRA, CNRS, Université Grenoble-Alpes, Grenoble, France. E-mail: [email protected] b Laboratoire des Technologies de la Microélectronique, CNRS, CEA, Université Grenoble-Alpes, Grenoble, France c Unité de Thérapie Cellulaire et Centre d'Investigation Clinique en Biothérapies, Hôpital Saint Louis, Institut Universitaire d'Hematologie, UMRS1160, INSERM, AP-HP, Université Paris Diderot, Paris, France d Laboratoire d'Electronique et de Technologie de l'Information, CEA, Université Grenoble-Alpes, Grenoble, France.
Hematopoietic stem cells (HSCs) are the most commonly used cell type in cell-based therapy. However, the investigation of their behaviour in vitro has been limited by the difficulty of monitoring these non-adherent cells under classical culture conditions. Indeed, fluid flow moves cells away from the video- recording position and prevents single cell tracking over long periods of time. Here we describe a large array of 2D no-flow chambers allowing the monitoring of single HSCs for several days. The chamber design has been optimized to facilitate manufacturing and routine use. The chip contains a single inlet and 800 chambers. The chamber medium can be renewed by diffusion within a few minutes. This allowed us to stain live human HSCs with fluorescent primary antibodies in order to reveal their stage in the haematopoiesis differentiation pathway. Thus we were able to correlate human HSCs' growth rate, polarization and migration to their differentiation stage.
… “a 4-wavelength LED excitation device (CoolLED pE-2 collimator, Life Sciences and Analytical), and”…
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pE-2: A repeatable, controllable modular system with 20 different LED peaks. Instant on/off and intensity (0-100%) control.
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