Hematopoietic Stem Cell Protocols / Edition 2

by Kevin D. Bunting

Overview

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 Prools 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 Prools thus aims to provide timely prools 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.

Product Details

ISBN-13:	9781588298683
Publisher:	Springer-Verlag New York, LLC
Publication date:	01/31/2008
Series:	Methods in Molecular Biology , #430
Edition description:	2nd ed. 2008
Pages:	312
Product dimensions:	6.10(w) x 9.25(h) x 0.04(d)

Table of 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 Prool for Bacterial Artificial Chromosome Recombineering to Study Essential Genes in Stem Cells.- Bioluminescence Imaging of Hematopoietic Stem Cell Repopulation in Murine Models.

Editorial Reviews

Reviewer: Bruce A. Fenderson, PhD (Thomas Jefferson University)
Description: Hematopoiesis is a complex developmental process characterized by stem cell renewal, lineage formation, and differentiation. This book, part of the user-friendly Methods in Molecular Biology series, provides detailed protocols for the identification, isolation, genetic manipulation, and differentiation of hematopoietic stem cells. These progenitor cells can be obtained from cultured embryonic stem (ES) cells, fetal tissues, and adult bone marrow. The book includes 20 chapters on topics ranging from stem cell homing to hematopoietic development of human ES cells. The primary focus is on cellular and developmental biology in human and mouse model systems.
Purpose: According to the editor, the aim of this book is to provide an "essential technical resource" for laboratory-based researchers interested in clinical applications of hematopoietic stem cells. His stated goal is to provide "updated approaches for the combination of phenotypic and functional analyses."
Audience: The book is written for basic science and clinical researchers interested in stem cell biology, hematology, and transplantation medicine. Biomedical scientists and clinicians interested in immunology, oncology, and regenerative medicine will also appreciate the wealth of information and exciting new ideas presented in this concise, hardcover book.
Features: Five major sections present an overview and information on stem cell enrichment, in vitro assays, transplantation assays, and genetic modification of hematopoietic stem cells. Each chapter includes a summary, introduction, list of essential materials and equipment, and step-by-step instructions for proven laboratory techniques. The authors present examples of results from their laboratories, such as protocols for flow cytometric analysis of mixed stem cell populations. Numerous line drawings, figures, and tables highlight key laboratory findings. A few figures are presented in color. The popular "Notes" section at the end of each chapter provides practical tips for achieving success in the laboratory. Each chapter concludes with a list of key references. The authors are experts in this clinically relevant and rapidly advancing field.
Assessment: The authors assume that readers are familiar with the basic biology of hematopoiesis, as well as current methods for cell sorting. The book builds on basic methods published in the first edition. In this second edition, the editor and authors highlight examples of cutting-edge research and emphasize new technology. Advances in stem cell biology will provide clinicians with new approaches to treating patients with congenital and acquired hematopathologies. The protocols in this book provide useful tools that will stimulate basic research.

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