Design of a 2D no-flow chamber to monitor hematopoietic stem cells.

Théo Cambier (1) , Thibault Honegger (2) , Valérie Vanneaux (3) , Jean Berthier (4) , David Peyrade (2) , Laurent Blanchoin (1) , Jerome Larghero (3) , Manuel Théry (1)

1 LPCV - Laboratoire de physiologie cellulaire végétale
2 LTM - Laboratoire des technologies de la microélectronique
3 (CIC-BT 301) - CIC - Biotherapie - Saint Louis
4 CEA-LETI - Commissariat à l'énergie atomique et aux énergies alternatives - Laboratoire d'Electronique et de Technologie de l'Information

Hematopoietic stem cells (HSCs) are the most commonly used cell type in cell-based therapy. However, the investigation of their behavior 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 hematopoiesis differentiation pathway. Thus we were able to correlate human HSCs' growth rate, polarization and migration to their differentiation stage.

Keywords: Technique; no flow chamber design; human; blood; stem cells

Domains: Life Sciences [q-bio]; Cellular Biology

Théo Cambier, Thibault Honegger, Valérie Vanneaux, Jean Berthier, David Peyrade, et al.. Design of a 2D no-flow chamber to monitor hematopoietic stem cells.. Lab on a Chip, 2015, 15 (1), pp.77-85. ⟨10.1039/c4lc00807c⟩. ⟨hal-01141132⟩