title: Marker-free detection of progenitor cell differentiation by analysis of Brownian motion in micro-wells.
abstract: The kinetics of stem and progenitor cell differentiation at the single-cell level provides essential clues to the complexity of the underlying decision-making circuits. In many hematopoietic progenitor cells, differentiation is accompanied by the expression of lineage-specific markers and by a transition from a non-adherent to an adherent state. Here, using the granulocyte-macrophage progenitor (GMP) as a model, we introduce a label-free approach that allows one to follow the course of this transition in hundreds of single cells in parallel. We trap single cells in patterned arrays of micro-wells and use phase-contrast time-lapse movies to distinguish non-adherent from adherent cells by an analysis of Brownian motion. This approach allowed us to observe the kinetics of induced differentiation of primary bone-marrow-derived GMPs into macrophages. The time lapse started 2 hours after addition of the cytokine M-CSF, and nearly 80% of the population had accomplished the transition within the first 20 h. The analysis of Brownian motion proved to be a sensitive and robust tool for monitoring the transition, and thus provides a high-throughput method for the study of cell differentiation at the single-cell level.
authors.affiliation: Faculty of Physics and Center for NanoScience, Ludwig-Maximilians-University, Munich, Germany. raedler@lmu.de.
authors.forename: Farzad
authors.initials: F
authors.lastname: Sekhavati
authors.forename: Max
authors.initials: M
authors.lastname: Endele
authors.forename: Susanne
authors.initials: S
authors.lastname: Rappl
authors.forename: Anna-Kristina
authors.initials: AK
authors.lastname: Marel
authors.forename: Timm
authors.initials: T
authors.lastname: Schroeder
authors.forename: Joachim O
authors.initials: JO
authors.lastname: Rädler
mesh_terms: D000818:Animals; D055592:Biophysical Phenomena; D002448:Cell Adhesion; D002454:Cell Differentiation; D019070:Cell Lineage; D004867:Equipment Design; D055014:Granulocyte-Macrophage Progenitor Cells / Q000166:cytology* / Q000502:physiology; D056656:Lab-On-A-Chip Devices; D051379:Mice; D008858:Microscopy, Phase-Contrast; D009038:Motion; D059010:Single-Cell Analysis / Q000295:instrumentation; D059008:Time-Lapse Imaging / Q000295:instrumentation