Hematopoietic Stem Cells: Methods and Protocols

by Louis M. Pelus (Editor), Jonathan Hoggatt (Editor)

Overview

This detailed volume collects updates on the technical advances in hematopoietic stem cell research and incorporates new techniques focused on the molecular/genetic, cellular, and whole organism levels. Exploring methods that apply stress to hematopoiesis, the book also contains chapters focused on better understanding the role of hematopoietic niches and their cellular components, as well as in vivo models that test and quantitate stem cell function and are key to further development of therapeutic applications. 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 and readily reproducible laboratory prools, and tips on troubleshooting and avoiding known pitfalls.

Authoritative and up-to-date, Hematopoietic Stem Cells: Methods and Prools serves as a valued addition to laboratories focused on understanding hematopoietic stem cell biology and the therapeutic advances that can be derived from it.

Product Details

ISBN-13:	9781071626818
Publisher:	Springer US
Publication date:	10/19/2023
Series:	Methods in Molecular Biology , #2567
Edition description:	1st ed. 2023
Pages:	282
Product dimensions:	7.01(w) x 10.00(h) x (d)

Table of Contents

The Factory of Blood Production: Hematopoietic Stem Cells.- Detection of DNA Damage in Hematopoietic Stem Cells.- Evaluating Histone Acetylation in Mouse Hematopoietic Stem and Progenitor Cells Using Chromatin Immunoprecipitation.- CRISPR Gene Editing of Hematopoietic Stem and Progenitor Cells.- Lentiviral Transduction of Nonhuman Primate Hematopoietic Stem and Progenitor Cells.- Identification of Nonhuman Primate Hematopoietic Stem and Progenitor Cells.- Isolation and Characterization of Fetal Liver Hematopoietic Stem Cells.- Utilizing CyTOF to Examine Hematopoietic Stem and Progenitor Phenotype.- Hematopoietic Stem Cell Identification Post-Radiation.- Intravital Microscopy for Hematopoietic Studies.- Laser Micromachining of Bone as a Tool for Studying Bone Marrow Biology.- MSC and HSPC Co-Culture: Mimicking Ex Vivo Bone Marrow Niche.- Isolation of Thymus Stromal Cells from Human and Murine Tissue.- Experimental Models of Mouse and Human Hematopoietic Stem Cell Transplantation.- Hematopoietic Stem and Progenitor Cell Identification and Transplantation in Zebrafish.- Establishing a Murine Model of the Hematopoietic Acute Radiation Syndrome.- Purinergic Signaling and Its Role in Mobilization of Bone Marrow Stem Cells.

Editorial Reviews

Reviewer: Omer Iqbal, MD, FACC, FESC (Loyola University Medical Center)
Description: Given rapid technical advances in hematopoietic stem cell research, it is now possible to use techniques focused on the molecular/genetic, cellular, and whole organism levels. Following the cloning of hematopoietic hormones, and development of FACS machines for analysis and selection of single cells, there was development of monoclonal antibodies, unique fluorochromes, and flow cytometry, which enabled these FACS machines to quantify stem cell and progenitor cell populations and signaling pathways in these cells. The book also provides methods that apply stress to hematopoiesis to better understand stem cell biology that would drive future advancements. The book also covers in vivo models that test and quantitate stem cell function, paving the way for therapeutic applications.
Purpose: The purpose of this book is to serve a valued addition to all laboratories focused on understanding hematopoietic stem cell biology and the therapeutic advances that can be derived. These are very worthy objectives and, hence, the book is very much needed. The book does meet the authors' objectives as it provides chapters focused on better understanding the role of hematopoietic niches and their cellular components in addition to the in vivo models that test and quantitate stem cell function that may lead to therapeutic applications.
Audience: The book is intended for those inexperienced or new to the field who apply these methods in their research, taking advantage of the guidance and discussions of the expert contributors. I absolutely agree with the authors and hope this book will also be useful for students, researchers, and hematologists interested in learning more about hematopoietic stem cell research. The book does meet the needs of the intended audience as it provides greater understanding of hematopoiesis and cell function. The authors are credible authorities in this field based on their credentials and lists of publications.
Features: The book is divided into five parts. Part I provides the overview and a chapter on the factory of blood production of hematopoietic stem cells. Part II covers the molecular modification and evolution processes and is subdivided into four interesting chapters discussing the CRISPR gene editing of hematopoietic stem (HSC) and progenitor cells (HPC), detection of DNA damage in hematopoietic stem cells, and lentiviral transduction of nonhuman primate hematopoietic stem and progenitor cells. Part III of the book covers cellular isolation and characterization and is subdivided into four chapters discussing the identification of nonhuman primate hematopoietic stem and progenitor cells and isolation and characterization of fetal liver hematopoietic stem cells. Another interesting chapter describes the single cell assay utilizing mass Cytometry by Time of Flight (CyTOF) technology to determine differences in expression profile within hematopoietic stem cells, using changes in HSC populations due to gender and aging. There is a chapter outlining techniques for HSC identification and analysis from mouse bone marrow post-radiation. Part IV of the book covers the hematopoietic niche and accessory cells and is subdivided into four chapters discussing the protocol and technique to visualize transplanted HSC in the mouse calvarium, using confocal and two-photon intravital microscopy. Another chapter provides a protocol for installing a laser-micromachining capability for users with an existing multiphoton microscope. Another interesting chapter on mesenchymal stromal cells (MSC) and hematopoietic stem/progenitor cells (HSPC) co-culture assay that can be used for ex vivo expansion of HSPC. The fourth chapter describes the methodology of harvesting thymus stromal cells from human and murine tissue for downstream applications such as flow cytometric analysis and single cell RNA sequencing. Part V of the book covers transplantation and in vivo models and is subdivided into four chapters describing experimental models of mouse and human HSC transplantation, and for assessment of donor cells collected and processed in hypoxic versus ambient air. Another chapter deals with the methods needed to identify, isolate and transplant HSPCs in zebrafish. One chapter is dedicated to establishing a murine model of the hematopoietic acute radiation syndrome. The last chapter in this book covers the role of purinergic signaling and extracellular nucleotides in regulating this process and reviews experimental strategies to study their involvement in mobilization of various types of stem cells that reside in the murine bone marrow. The book has several illustrations, which clarify understanding of the subject presented. Expanding the index may have enabled much easier location of the subject matter.
Assessment: This is an excellent book on hematopoietic stem cells. It discusses new techniques focused on the molecular, cellular, and whole organism levels as well as updated methods and protocols that apply stress to hematopoiesis, a major pathway to understand HSC biology. The book also presents information on in vivo models to test and quantitate stem cell function, which is crucial to development of therapeutic applications. The intended audience of students, hematologists, and inexperienced and experienced researchers will find this book very interesting and it will help drive future research in exciting new directions.

Doody's Review Service