Hematopoietic Stem Cell Protocols

by Kevin D. Bunting (Editor), Cheng-Kui Qu (Editor)

Overview

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.

Product Details

ISBN-13:	9781493947577
Publisher:	Springer New York
Publication date:	09/17/2016
Series:	Methods in Molecular Biology , #1185
Edition description:	Softcover reprint of the original 3rd ed. 2014
Pages:	364
Product dimensions:	7.01(w) x 10.00(h) x (d)

Table of Contents

Preface     v
Contributors     xi
Overview
The Expanding Tool Kit for Hematopoietic Stem Cell Research   William Tse   Kevin D. Bunting     3
Stem Cell Enrichment and Analysis
Isolation of Quiescent Murine Hematopoietic Stem Cells by Homing Properties   Tarja A. Juopperi   Saul J. Sharkis     21
Mobilization of Hematopoietic Stem and Progenitor Cells in Mice   Simon N. Robinson   Ronald P. van Os     31
Transmigration of Human CD34[superscript +] Cells   Seiji Fukuda   Louis M. Pelus     55
Flow Cytometry-Based Cell Cycle Measurement of Mouse Hematopoietic Stem and Progenitor Cells   Hongmei Shen   Matthew Boyer   Tao Cheng     77
Analysis of Apoptosis in Hematopoietic Stem Cells by Flow Cytometry   William G. Kerr     87
In Vitro Assays and Differentiation
In Vitro Hematopoietic Differentiation of Murine Embryonic Stem Cells   Jinhua Shen   Cheng-Kui Qu     103
Hematopoietic Development of Human Embryonic Stem Cells in Culture   Xinghui Tian   Dan S. Kaufman     119
In Vitro Human T Cell Development Directed by Notch-Ligand Interactions   Geneve Awong   Ross N. La Motte-Mohs   Juan Carlos   Zuniga-Pflucker     135
In Vitro Assays for Cobblestone Area-Forming Cells, LTC-IC, and CFU-C   Ronald P. van Os   Bertien Dethmers-Ausema   Gerald de Haan     143
Transplantation Assays for Mouse and Human HSC
Hematopoietic Stem Cell Transplant in Mice by Intra-Femoral Injection   Yuxia Zhan   Yi Zhao     161
Hematopoietic Stem Cell Transplant into Non-Myeloablated W/W[superscript v] Mice to Detect Steady-State Engraftment Defects   Zhengqi Wang   Kevin D. Bunting     171
Isolation and Functional Characterization of Side Population Stem Cells   Jonathan B. Johnnidis   Fernando D. Camargo     183
Transplantation of Chimeric Fetal Liver to Study Hematopoiesis   Sigrid Eckardt   K. John McLaughlin     195
Immunodeficient Mouse Models to Study Human Stem Cell-Mediated Tissue Repair   Ping Zhou   Sarah Hohm   Ben Capoccia   Louisa Wirthlin   David Hess   Dan Link   Jan Nolta     213
Genetic Modification of HSCs and Imaging Engraftment
Retroviral Transduction of Murine Hematopoietic Stem Cells   Peter Haviernik   Yi Zhang   Kevin D. Bunting     229
Lentiviral Gene Transduction of Mouse and Human Stem Cells   Zhaohui Ye   Xiaobing Yu   Linzhao Cheng     243
Retroviral Integration Site Analysis in Hematopoietic Stem Cells   Olga S. Kustikova   Christopher Baum   Boris Fehse     255
A Detailed Protocol for Bacterial Artificial Chromosome Recombineering to Study Essential Genes in Stem Cells   Andriy Tsyrulnyk   Richard Moriggl     269
Bioluminescence Imaging of Hematopoietic Stem Cell Repopulation in Murine Models   Yuan Lin   Joe Molter   Zhenghong Lee   Stanton L. Gerson     295
Index     307

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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