- Email: [email protected]
Hematopoietic Stem Cell Protocols: Hematopoietic stem cell analysis ‘step by step’
464
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chapter. The author list includes many very familiar names, and thus it is not surprising to find a lot of very good material in the book. However, the coherent plan promised by the table of contents is not always realized and there is little sense of strong editorial direction. Thus, while the volume is more than a collection of current reviews, it does not quite hold together as a textbook. For example, quite different concepts of what constitutes an overview were taken by the authors of the four opening chapters, entitled ‘overview of the bacterial/fungal/ parasitic/viral pathogens’; the bacterial chapter focusing on a few virulence genes seemed out of place. Another consequence of this editorial approach is that there is a lot of redundancy between chapters. For example, almost every parasite chapter seemed to have a discussion of the Th1/Th2 paradigm. Although this would be confusing in a textbook, it does provide interesting differing perspectives on a given topic. By contrast, some topics (such as complement and NK cells) are not explicitly covered but nevertheless crop up in various different chapters, with the result that most things are discussed somewhere. Inevitably, I found some of the chapters more useful than others. Throughout the book the most enjoyable chapters for me were those on parasites and fungi, no doubt because I am least familiar with them. I came away with a real appreciation of how the body defends itself against these ubiquitous organisms, as well as a renewed appreciation for the complement pathway and Th2 cells. In summary, anyone with a solid background in immunology and with at least basic bacteriology, who wants to consolidate and broaden their understanding of how mammalian hosts defend themselves against pathogens, could profitably read this book. I see it as a useful resource for infectious disease or immunologist clinicians and for college level teachers. My feeling is that this volume will be most useful when read in its entirety and anyone who does so will be rewarded with a deeper understanding of host defense. Ann Hill Dept of Molecular Microbiology & Immunology, Oregon Health Sciences University, 3181 SW Sam Jackson Park Road, Portland, OR 97201-3098, USA. e-mail: [email protected] http://immunology.trends.com
TRENDS in Immunology Vol.23 No.9 September 2002
Hematopoietic stem cell analysis ‘step by step’ Hematopoietic Stem Cell Protocols (Methods in Molecular Medicine) edited by Christopher A. Klug and Craig T. Jordan. Humana Press, 2001.US$99.50 (hdbk) (xi + 332 pages) ISBN 0 896 03812 2
The past decade has produced remarkable advances in our ability to characterize the various cell lineages of human hematopoietic stem cells (HSCs). HSCs are self-renewing progenitors that give rise to all lineages of blood cells. They are found in all hematopoietic organs, from para-aortic mesoderm and yolk sac in fetuses to the bone marrow, blood and spleens of adults. Our knowledge of HSCs can be applied successfully in clinical practice. The most obvious immediate benefit is the possibility of increasing the chemotherapeutic dose in cancer and hematological malignancies by way of hematopoietic stem cell transplantation. This technique has potential in many clinical settings, including the treatment of other non-malignant diseases. Medical knowledge is constantly renewed. This is particularly true for hematopoietic stem-cell transplantation, hematopoietic gene therapy, solid organ transplantation and somatic tissue regeneration. The technology involved is developing rapidly and in this context the selection of stem cells and their characterization and expansion in vitro are vital processes. Perhaps the most exciting potential of cell therapy is our ability to manipulate cells by modifying their physiological properties before returning them to a patient. Cell therapy is evolving quickly, drawing on cell biology, molecular biology, virology, immunology and cell quantification techniques and also on biomedical engineering. These advances require well defined, reproducible and firmly established laboratory methods for investigating HSCs. The hematopoietic system comprises a concentrated series of stem and transit progenitor cell compartments of progressively restricted potentiality and proliferative capacity. In this setting, Hematopoietic Stem Cell Protocols, edited by Christopher A. Klug and Craig T. Jordan, is an essential handbook for novice and
experienced investigators alike, which gives a wide variety of step-by-step instructions for the study of mouse and human HSCs of both embryonic and adult origin. The 20 chapters present techniques for stem cell analysis, some of which have become standardized over the past few years. Many chapters contain contributions by the researchers who first developed and introduced the procedures. Each chapter begins with a short general introduction referring to most of the main publications in the field, then the major part precisely reports the materials and methods, giving technical laboratory details or even ‘home-made cooking secrets’. Methods are described in the following areas: aorta–gonad–mesonephros and yolk sac HSCs; isolation of mouse HSCs; flow cytometric analysis and immunoselection of HSCs; purification of human HSCs by flow cytometry; cycling and turnover of HSC; cell-cycle analysis of primitive stem cells; hematopoietic colony-forming cells; long-term culture-initiating cell (LTC-IC) assays for human and murine cells; assays for cobblestone area-forming cells; colony forming unit on the spleen (CFU-S) assays; T-cell progenitor activity and competitive repopulation assays for HSCs in the NOD and SCID models. Protocols for stem cell expansion and the hematopoietic maturation of ES cells are included, as are detailed methods for the genetic modification of stem cells, for example, retroviral infection of murine HSCs, retroviral transduction of purified HSCs, retroviral-mediated transduction and HIVbased vectors. The last section deals with gene expression in HSCs and the use of twodimensional gene-expression fingerprinting. This is a comprehensive compendium of practical techniques, a state-of-the-art handbook. It will be a useful tool for teams involved in either basic hematopoiesis research or progenitoror stem cell quantification for cell therapy protocols in the context of patient care. Thanks to a multidisciplinary approach, from fundamental to practical aspects, this book reports advances made in the field of hematopoiesis, which could subsequently be applied for diagnostic and therapeutic purposes. Luc Douay Laboratoire d’Hématologie, Hôpital Armand Trousseau, 26 Avenue du Docteur Netter, 75012 Paris, France. e-mail: [email protected]
1471-4906/02/$ – see front matter © 2002 Elsevier Science Ltd. All rights reserved.
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chapter. The author list includes many very familiar names, and thus it is not surprising to find a lot of very good material in the book. However, the coherent plan promised by the table of contents is not always realized and there is little sense of strong editorial direction. Thus, while the volume is more than a collection of current reviews, it does not quite hold together as a textbook. For example, quite different concepts of what constitutes an overview were taken by the authors of the four opening chapters, entitled ‘overview of the bacterial/fungal/ parasitic/viral pathogens’; the bacterial chapter focusing on a few virulence genes seemed out of place. Another consequence of this editorial approach is that there is a lot of redundancy between chapters. For example, almost every parasite chapter seemed to have a discussion of the Th1/Th2 paradigm. Although this would be confusing in a textbook, it does provide interesting differing perspectives on a given topic. By contrast, some topics (such as complement and NK cells) are not explicitly covered but nevertheless crop up in various different chapters, with the result that most things are discussed somewhere. Inevitably, I found some of the chapters more useful than others. Throughout the book the most enjoyable chapters for me were those on parasites and fungi, no doubt because I am least familiar with them. I came away with a real appreciation of how the body defends itself against these ubiquitous organisms, as well as a renewed appreciation for the complement pathway and Th2 cells. In summary, anyone with a solid background in immunology and with at least basic bacteriology, who wants to consolidate and broaden their understanding of how mammalian hosts defend themselves against pathogens, could profitably read this book. I see it as a useful resource for infectious disease or immunologist clinicians and for college level teachers. My feeling is that this volume will be most useful when read in its entirety and anyone who does so will be rewarded with a deeper understanding of host defense. Ann Hill Dept of Molecular Microbiology & Immunology, Oregon Health Sciences University, 3181 SW Sam Jackson Park Road, Portland, OR 97201-3098, USA. e-mail: [email protected] http://immunology.trends.com
TRENDS in Immunology Vol.23 No.9 September 2002
Hematopoietic stem cell analysis ‘step by step’ Hematopoietic Stem Cell Protocols (Methods in Molecular Medicine) edited by Christopher A. Klug and Craig T. Jordan. Humana Press, 2001.US$99.50 (hdbk) (xi + 332 pages) ISBN 0 896 03812 2
The past decade has produced remarkable advances in our ability to characterize the various cell lineages of human hematopoietic stem cells (HSCs). HSCs are self-renewing progenitors that give rise to all lineages of blood cells. They are found in all hematopoietic organs, from para-aortic mesoderm and yolk sac in fetuses to the bone marrow, blood and spleens of adults. Our knowledge of HSCs can be applied successfully in clinical practice. The most obvious immediate benefit is the possibility of increasing the chemotherapeutic dose in cancer and hematological malignancies by way of hematopoietic stem cell transplantation. This technique has potential in many clinical settings, including the treatment of other non-malignant diseases. Medical knowledge is constantly renewed. This is particularly true for hematopoietic stem-cell transplantation, hematopoietic gene therapy, solid organ transplantation and somatic tissue regeneration. The technology involved is developing rapidly and in this context the selection of stem cells and their characterization and expansion in vitro are vital processes. Perhaps the most exciting potential of cell therapy is our ability to manipulate cells by modifying their physiological properties before returning them to a patient. Cell therapy is evolving quickly, drawing on cell biology, molecular biology, virology, immunology and cell quantification techniques and also on biomedical engineering. These advances require well defined, reproducible and firmly established laboratory methods for investigating HSCs. The hematopoietic system comprises a concentrated series of stem and transit progenitor cell compartments of progressively restricted potentiality and proliferative capacity. In this setting, Hematopoietic Stem Cell Protocols, edited by Christopher A. Klug and Craig T. Jordan, is an essential handbook for novice and
experienced investigators alike, which gives a wide variety of step-by-step instructions for the study of mouse and human HSCs of both embryonic and adult origin. The 20 chapters present techniques for stem cell analysis, some of which have become standardized over the past few years. Many chapters contain contributions by the researchers who first developed and introduced the procedures. Each chapter begins with a short general introduction referring to most of the main publications in the field, then the major part precisely reports the materials and methods, giving technical laboratory details or even ‘home-made cooking secrets’. Methods are described in the following areas: aorta–gonad–mesonephros and yolk sac HSCs; isolation of mouse HSCs; flow cytometric analysis and immunoselection of HSCs; purification of human HSCs by flow cytometry; cycling and turnover of HSC; cell-cycle analysis of primitive stem cells; hematopoietic colony-forming cells; long-term culture-initiating cell (LTC-IC) assays for human and murine cells; assays for cobblestone area-forming cells; colony forming unit on the spleen (CFU-S) assays; T-cell progenitor activity and competitive repopulation assays for HSCs in the NOD and SCID models. Protocols for stem cell expansion and the hematopoietic maturation of ES cells are included, as are detailed methods for the genetic modification of stem cells, for example, retroviral infection of murine HSCs, retroviral transduction of purified HSCs, retroviral-mediated transduction and HIVbased vectors. The last section deals with gene expression in HSCs and the use of twodimensional gene-expression fingerprinting. This is a comprehensive compendium of practical techniques, a state-of-the-art handbook. It will be a useful tool for teams involved in either basic hematopoiesis research or progenitoror stem cell quantification for cell therapy protocols in the context of patient care. Thanks to a multidisciplinary approach, from fundamental to practical aspects, this book reports advances made in the field of hematopoiesis, which could subsequently be applied for diagnostic and therapeutic purposes. Luc Douay Laboratoire d’Hématologie, Hôpital Armand Trousseau, 26 Avenue du Docteur Netter, 75012 Paris, France. e-mail: [email protected]
1471-4906/02/$ – see front matter © 2002 Elsevier Science Ltd. All rights reserved.