Stem Cell Mobilization Methods and Protocols

This book provides detailed state-of-the-art protocols for analyzing individual aspects of stem cell mobilization at the molecular and cellular level with contributions encompassing the most recent technological developments. Specifically, the volume covers cell adhesion and migration assays in three-dimensional models, imaging techniques for the determination of blood flow or hypoxic status, innovative biomechanical studies of stem cells, and assays to determine the involvement of proteases or complement factors, as well as other key topics. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. 

1. Evolution of Peripheral Blood Stem Cell Transplantation

            Anthony D. Ho

 

2. In Vitro Dynamic Phenotyping for Testing Novel Mobilizing Agents

            Motomu Tanaka

 

3. Colony Formation: An Assay of Hematopoietic Progenitor Cells

            Romy Kronstein-Wiedemann and Torsten Tonn

 

4. Mobilization and Collection of Peripheral Blood Stem Cells in Adults: Focus on Timing and Benchmarking

            Katharina Kriegsmann and Patrick Wuchter

 

5. Transmigration Assays for the Determination of Molecular Interactions between Hematopoietic Stem Cells and Niche Cells

            Reinhard Henschler and Rudolf Richter

 

6. Microfluidic Shear Force Assay to Determine Cell Adhesion Forces

            Julia Hümmer, Julian Koc, Axel Rosenhahn, and Cornelia Lee-Thedieck

 

7. A Microcavity Array-Based 3D-Model System of the Hematopoietic Stem Cell Niche

            Eric Gottwald, Cordula Nies, Patrick Wuchter, Rainer Saffrich, Roman Truckenmüller, and Stefan Giselbrecht

 

8. Migration Assay for Leukemic Cells in a 3D Matrix toward a Chemoattractant

            Sabrina Zippel, Annamarija Raic, and Cornelia Lee-Thedieck

 

9. Intravital Imaging of Blood Flow and HSPC Homing in Bone Marrow Microvessels

            Jonas Stewen and Maria Gabriele Bixel

 

10. Assessing Cellular Hypoxic Status In Situ within the Bone Marrow Microenvironment

            Ute Suessbier and César Nombela-Arrieta

 

11. Analysis of Biomechanical Properties of Hematopoietic Stem and Progenitor Cells Using Real-Time Fluorescence and Deformability Cytometry

            Angela Jacobi, Philipp Rosendahl, Martin Kräter, Marta Urbanska, Maik Herbig, and Jochen Guck

 

12. Assessment of Proteolytic Activities in the Bone Marrow Microenvironment

            Andreas Maurer, Gerd Klein, and Nicole D. Staudt

 

13. Analysis of the Complement Cascade Activation during Mobilization of Hematopoietic Stem/Progenitor Cells

            Anna I. Grabowska and Jakub M. Hawryluk

 

14. A Freezing Protocol for Hematopoietic Stem Cells

            Petra Pavel and Sascha Laier

 

15. Assessment of Young and Aged Hematopoietic Stem Cell Activity by Competitive Serial Transplantation Assays

            Yu Wei Zhang and Nina Cabezas Wallscheid

 

16. Zebrafish Xenografts for the In Vivo Analysis of Healthy and Malignant Human Hematopoietic Cells

            Martina Konantz, Joëlle S. Müller, and Claudia Lengerke

 

17. Statistical and Mathematical Modeling of Spatiotemporal Dynamics of Stem Cells

            Walter de Back, Thomas Zerjatke, and Ingo Roeder