Updating and building upon previous editions, Hematopoietic Stem Cell Protocols, Third Edition provides up-to-date protocols from leading stem cell researchers. This in-depth volume presents a clear view of the landscape of assays available to the stem cell researcher working in the growing hematopoietic stem cell (HSC) field. A robust and active field, it is supported by an abundance of innovative mouse models and molecular tools for analysis of phenotypes and functions in mouse and human cells. Understanding more about hematopoietic stem cell biology is integral if these versatile cells are to be applied effectively to treat and cure a wide range of blood diseases. An introductory chapter puts the major contributions of the book into the proper perspective. Written in the successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible protocols, and notes on troubleshooting and avoiding known pitfalls.

Essential for the laboratory-based researcher, Hematopoietic Stem Cell Protocols, Third Edition is a much needed technical resource in the critically important field of hematopoietic stem cell investigation.

Includes cutting-edge methods and protocols Provides step-by-step detail essential for reproducible results Contains key notes and implementation advice from the experts Includes supplementary material: sn.pub/extras

Inhalt

The Hematopoietic Stem Cell Landscape.- Investigating the Interaction Between Hematopoietic Stem Cells and Their Niche During Embryonic Development: Optimizing the Isolation of Fetal and Newborn Stem Cells From Liver, Spleen, and Bone Marrow.- Single-Cell PCR Profiling of Gene Expression in Hematopoiesis.- Hematopoietic Stem and Progenitor Cell Mobilization in Mice.- Cell Cycle Measurement of Mouse Hematopoietic Stem/Progenitor Cells.- Flow Cytometry Analysis of Signaling and Apoptosis in Hematopoietic Stem Cells.- Gene Expression Profiling of Hematopoietic Stem Cells (HSC).- Measuring MicroRNA Expression in Mouse Hematopoietic Stem Cells.- DNA Methylation Profiling of Hematopoietic Stem Cells.- Metabolic Characterization of Hematopoietic Stem Cells.- Nano Proteomic Assays on Hematopoietic Stem Cells.- Hematopoietic Differentiation of Pluripotent Stem Cells in Culture.- Ex vivo Assays to Study Self-Renewal, Long-Term Expansion and Leukemic Transformation of Genetically Modified Human Hematopoietic and Patient-Derived Leukemic Stem Cells.- Ex vivo Expansion of Murine and Human Hematopoietic Stem Cells.- Using Microfluidics to Investigate Hematopoietic Stem Cell and Micro niche Interactions at the Single Cell Level.- Five Lineage Clonal Analysis of Hematopoietic Stem/Progenitor Cells.- Intravital Imaging of Hematopoietic Stem Cells in the Mouse Skull.- Immune Deficient Mouse Model for Human Hematopoietic Stem Cell Engraftment and Immune System Development.- Homing and Migration Assays of Hematopoietic Stem/Progenitor Cells.- Retroviral Transduction of Murine and Human Hematopoietic Progenitors and Stem Cells.- Lentiviral Gene Transduction of Mouse and Human Hematopoietic Stem Cells.- High Throughput Genomic Mapping of Vector Integration Sites in Gene Therapy Studies.- Barcoded Vector Libraries and Retro- or Lentiviral Barcoding of Hematopoietic Stem Cells.- Barcoded Vector Libraries and Retro- or Lentiviiral Barcoding of Hematopoietic StemCells.- Using Microfluidics to Investigate Hematopoietic Stem Cell and Micro niche Interactions at the Single Cell Level.- Five Lineage Clonal Analysis of Hematopoietic Stem/Progenitor Cells.- Intravital Imaging of Hematopoietic Stem Cells in the Mouse Skull.- Immune Deficient Mouse Model for Human Hematopoietic Stem Cell Engraftment and Immune System Development.- Homing and Migration Assays of Hematopoietic Stem/Progenitor Cells.- Retroviral Transduction of Murine and Human Hematopoietic Progenitors and Stem Cells.- Lentiviral Gene Transduction of Mouse and Human Hematopoietic Stem Cells.- High Throughput Genomic Mapping of Vector Integration Sites in Gene Therapy Studies.- Barcoded Vector Libraries and Retro- or Lentiviiral Barcoding of Hematopoietic Stem Cells.