Analysis of Biomechanical Properties of Hematopoietic Stem and Progenitor Cells Using Real-Time Fluorescence and Deformability Cytometry
Stem cell mechanics, determined predominantly by the cell’s cytoskeleton, plays an important role in different biological processes such as stem cell differentiation or migration. Several methods to measure mechanical properties of cells are currently available, but most of them are limited in the ability to screen large heterogeneous populations in a robust and efficient manner—a feature required for successful translational applications. With real-time fluorescence and deformability cytometry (RT-FDC), mechanical properties of cells in suspension can be screened continuously at rates of up to 1,000 cells/s—similar to conventional flow cytometers—which makes it a suitable method not only for basic research but also for a clinical setting. In parallel to mechanical characterization, RT-FDC allows to measure specific molecular markers using standard fluorescence labeling. In this chapter, we provide a detailed protocol for the characterization of hematopoietic stem and progenitor cells (HSPCs) in heterogeneous mobilized peripheral blood using RT-FDC and present a specific morpho-rheological fingerprint of HSPCs that allows to distinguish them from all other blood cell types.
Key wordsMechanical phenotyping Hematopoietic stem and progenitor cells Cell mechanics Microfluidics Flow cytometry
The authors would like to thank Prof. Martin Bornhäuser from the University Hospital Dresden for providing the patients material, Zellmechanik Dresden for providing materials for graphics, and the Microstructure Facility at the Center for Molecular and Cellular Bioengineering (CMCB) at Technische Universität Dresden (in part funded by the State of Saxony and the European Regional Development Fund) and Alejandro Riviera Prieto for help with the production of RT-DC chips. This work was financially supported by the Alexander von Humboldt-Stiftung (Alexander von Humboldt Professorship to J.G.) and the DKMS Mechthild Harf Research Grant (DKMS-SLS-MHG-2016-02 to A.J.).
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