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Full Description
The hematopoietic stem cell (HSC) field has rapidly grown in the past several years as new technologies have been developed and the older tried and true methods have been used in new ways. Major advances in the isolation of HSC and progenitor subsets have been sparked by the gene expression array technologies, leading to the identification of new markers for prospective isolation. The first edition of Hematopoietic Stem Cell Protocols is a very thorough resource covering historically the basic techniques that have been published extensively over many years. To put together, a follow-up to this excellent book has been a hard act to follow. The intent of this book is to be distinct and not duplicate but to build on the strong basic assays and include updated approaches for the combination of phenotypic and functional analyses. This second edition of Hematopoietic Stem Cell Protocols thus aims to provide timely protocols developed in the HSC field since the last comprehensive review in this series. The approaches described in this book should be readily useful for the basic science researcher, especially those familiar with the use of mouse genetic models. The chapters in this book are geared toward the laboratory-based researcher and the development of pre-clinical studies to apply HSCs toward treating blood diseases more effectively.
Contents
Overview.- The Expanding Tool Kit for Hematopoietic Stem Cell Research.- Stem Cell Enrichment and Analysis.- Isolation of Quiescent Murine Hematopoietic Stem Cells by Homing Properties.- Mobilization of Hematopoietic Stem and Progenitor Cells in Mice.- Transmigration of Human CD34+ Cells.- Flow Cytometry-Based Cell Cycle Measurement of Mouse Hematopoietic Stem and Progenitor Cells.- Analysis of Apoptosis in Hematopoietic Stem Cells by Flow Cytometry.- In Vitro Assays and Differentiation.- In Vitro Hematopoietic Differentiation of Murine Embryonic Stem Cells.- Hematopoietic Development of Human Embryonic Stem Cells in Culture.- In Vitro Human T Cell Development Directed by Notch-Ligand Interactions.- In Vitro Assays for Cobblestone Area-Forming Cells, LTC-IC, and CFU-C.- Transplantation Assays for Mouse and Human HSC.- Hematopoietic Stem Cell Transplant in Mice by Intra-Femoral Injection.- Hematopoietic Stem Cell Transplant into Non-Myeloablated W/W v Mice to Detect Steady-State Engraftment Defects.- Isolation and Functional Characterization of Side Population Stem Cells.- Transplantation of Chimeric Fetal Liver to Study Hematopoiesis.- Immunodeficient Mouse Models to Study Human Stem Cell-Mediated Tissue Repair.- Genetic Modification of HSCs and Imaging Engraftment.- Retroviral Transduction of Murine Hematopoietic Stem Cells.- Lentiviral Gene Transduction of Mouse and Human Stem Cells.- Retroviral Integration Site Analysis in Hematopoietic Stem Cells.- A Detailed Protocol for Bacterial Artificial Chromosome Recombineering to Study Essential Genes in Stem Cells.- Bioluminescence Imaging of Hematopoietic Stem Cell Repopulation in Murine Models.
