Protocols for Adult Stem Cells / Edition 1

by Irina M. Conboy, David V. Schaffer, Mary Helen Barcellos-Hoff, Song Li

Overview

Although embryonic stem cells currently enjoy the public limelight and show great pr- ise for cell based medical therapies, it is the adult stem cells which are responsible for the body’s natural ability to fght disease, heal and recover, or fail and succumb to various maladies. The study of mammalian adult stem cells has surged recently, most likely from a maturation of stem cell studies in the classical developmental model organisms and in hematopoeisis. All the tissues of the body examined so far are generated and regenerated from stem cells, it has been an important frst step to adapt or devise new methods to identify and obtain these cells in quantity and purity for further study. Culture techniques have been optimized for managing the growth and differentiation of stem cells in vitro; as some stem cells are pluripotent, often the method is to guide the fate of such cells among the possible differentiation fates. Much of this work, and that in the classical model org- isms, has helped defne the aspects of the stem cell environment or niche that are crucial for both growth and differentiation, and these studies have moved in vivo at increasingly higher resolution. Importantly, the in vivo niche is a current target for bioengineering the matrix and signaling factors. Herein, we present methods for studying six types of mammalian stem cells, m- mary, neural, mesenchymal, endothelial, dendritic, and muscle.

Product Details

ISBN-13:	9781607610625
Publisher:	Springer-Verlag New York, LLC
Publication date:	04/22/2010
Series:	Methods in Molecular Biology , #621
Edition description:	2010
Pages:	207
Product dimensions:	7.20(w) x 10.10(h) x 0.80(d)

Table of Contents

Preface v

Contributors ix

1 In Situ Analysis of Cell Populations: Long-Term Label-Retaining Cells Rodrigo Fernandez-Gonzalez Irineu Illa-Bochaca Dawne N. Shelton Bryan E. Welm Mary Helen Barcellos-Hoff Carlos Ortiz-de-Solorzano 1

2 Limiting-Dilution Transplantation Assays in Mammary Stem Cell Studies Irineu Illa-Bochaca Rodrigo Fernandez-Gonzalez Dawne N. Shelton Bryan E. Welm Carlos Ortiz-de-Solorzano Mary Helen Barcellos-Hoff 29

3 Use of Stem Cell Markers in Dissociated Mammary Populations Dawne N. Shelton Rodrigo Fernandez-Gonzalez Irineu Illa-Bochaca Carlos Ortiz-de-Solorzano Mary Helen Barcellos-Hoff Bryan E. Welm 49

4 Isolation of Adult Hippocampal Neural Progenitors Joseph Peltier Brandi K. Ormerod David V. Schaffer 57

5 In Vitro Culture and Analysis of Adult Hippocampal Neural Progenitors Joseph Peltier Smita Agrawal Matthew J. Robertson David V. Schaffer 65

6 In Vivo Analysis of Engrafted Adult Hippocampal Neural Progenitors Matthew J. Robertson Joseph Peltier David V. Schaffer 89

7 Viral Packaging and Transduction of Adult Hippocampal Neural Progenitors Joseph Peltier David V. Schaffer 103

8 Culture of Bone Marrow Mesenchymal Stem Cells on Engineered Matrix Jennifer S. Park Craig Hashi Song Li 117

9 Preparation and Analysis of Endothelial Progenitor Cells Associated with Angiogenesis Steve Khachi Yiqian Zhu Guo-Yuan Yang 139

10 Preparation of Adult Muscle Fiber-Associated Stem/Precursor Cells Michael J. Conboy Irina M. Conboy 149

11 Immuno-Analysis and FACS Sorting of Adult Muscle Fiber-Associated Stem/Precursor Cells Michael J. Conboy Massimiliano Cerletti Amy J. Wagers Irina M. Conboy 165

12 Altered Expression of NFκB in Ex Vivo Differentiated Dendritic Cells from the Aged Subjects: Implications in Immunotherapy Anshu Agrawal 175

13 Preparation of Collagen-Coated Gels that Maximize in Vitro Myogenesis of Stem Cells by Matching the Lateral Elasticity of in Vivo Muscle Tathagata Chaudhuri Florian Rehfeldt H. Lee Sweeney Dennis E. Discher 185

Index 203

Editorial Reviews

Reviewer: Bruce A. Fenderson, PhD (Thomas Jefferson University)
Description: Tissues in the body are populated with stem cells that are able to self-renew and differentiate to compensate for cell loss due to senescence and injury. This volume in the popular Methods in Molecular Biology series provides a snapshot of research techniques and protocols for identifying and culturing these amazing tissue-specific stem cells. The 13 chapters cover topics ranging from identification of mammary epithelial stem cells to the use of cultured dendritic cells for anticancer immunotherapy. The primary focus is on mammalian stem cell culture and modern techniques in cell biology.
Purpose: According to the editors, the goals of this volume are to make available "new methods to identify and obtain these [stem cells] in quantity and purity for further study." The editors and authors are keenly interested in defining "aspects of the stem cell environment or niche that are crucial for both growth and differentiation." Future applications of adult stem cell research focus on the restoration of organ function in patients with chronic disease.
Audience: The book is written for basic science and clinical researchers interested in stem cell biology, as well as mechanisms of carcinogenesis and malignancy. It also will be valued by biomedical scientists and clinicians interested in cytotechnology and biotechnology.
Features: This volume provides essential background information and detailed protocols for identifying, isolating, characterizing, and manipulating adult organ-specific stem cells. Each chapter contains an abstract, overview and introduction, step-by-step materials and methods, notes, and key references. The notes section provides helpful tips for troubleshooting and avoiding known pitfalls. The authors highlight results and provide examples of stem cell morphology in culture. This volume focuses on six types of stem cells, those for the mammary gland, connective tissue mesenchyme, vascular endothelium, dendritic cells, skeletal muscle, and central nervous system. Some of the figures are reproduced in color.
Assessment: A minor criticism is that the techniques used in this field are still primitive (e.g., fat pad injection assay). In addition, there is a lack of consensus on definitive stem cell markers. Stem cell culture remains an art more than a science. Nonetheless, this book will be valued by basic science and clinical researchers because it pulls together methods that are yielding insights into isolation of tissue-specific stem cells and their culture requirements. The topic is fascinating and clinically relevant. I enjoyed learning about antigen-priming dendritic cells.

Doody's Review Service

Reviewer: Bruce A. Fenderson, PhD (Thomas Jefferson University)
Description: Tissues in the body are populated with stem cells that are able to self-renew and differentiate to compensate for cell loss due to senescence and injury. This volume in the popular Methods in Molecular Biology series provides a snapshot of research techniques and protocols for identifying and culturing these amazing tissue-specific stem cells. The 13 chapters cover topics ranging from identification of mammary epithelial stem cells to the use of cultured dendritic cells for anticancer immunotherapy. The primary focus is on mammalian stem cell culture and modern techniques in cell biology.
Purpose: According to the editors, the goals of this volume are to make available "new methods to identify and obtain these [stem cells] in quantity and purity for further study." The editors and authors are keenly interested in defining "aspects of the stem cell environment or niche that are crucial for both growth and differentiation." Future applications of adult stem cell research focus on the restoration of organ function in patients with chronic disease.
Audience: The book is written for basic science and clinical researchers interested in stem cell biology, as well as mechanisms of carcinogenesis and malignancy. It also will be valued by biomedical scientists and clinicians interested in cytotechnology and biotechnology.
Features: This volume provides essential background information and detailed protocols for identifying, isolating, characterizing, and manipulating adult organ-specific stem cells. Each chapter contains an abstract, overview and introduction, step-by-step materials and methods, notes, and key references. The notes section provides helpful tips for troubleshooting and avoiding known pitfalls. The authors highlight results and provide examples of stem cell morphology in culture. This volume focuses on six types of stem cells, those for the mammary gland, connective tissue mesenchyme, vascular endothelium, dendritic cells, skeletal muscle, and central nervous system. Some of the figures are reproduced in color.
Assessment: A minor criticism is that the techniques used in this field are still primitive (e.g., fat pad injection assay). In addition, there is a lack of consensus on definitive stem cell markers. Stem cell culture remains an art more than a science. Nonetheless, this book will be valued by basic science and clinical researchers because it pulls together methods that are yielding insights into isolation of tissue-specific stem cells and their culture requirements. The topic is fascinating and clinically relevant. I enjoyed learning about antigen-priming dendritic cells.

Doody Reviews

From the reviews:

“This volume in the popular Methods in Molecular Biology series provides a snapshot of research techniques and prools for identifying and culturing these amazing tissue-specific stem cells. … The book is written for basic science and clinical researchers interested in stem cell biology, as well as mechanisms of carcinogenesis and malignancy. It also will be valued by biomedical scientists and clinicians interested in cytotechnology and biotechnology. … The topic is fascinating and clinically relevant. I enjoyed learning about antigen-priming dendritic cells.” (Bruce A. Fenderson, Doody’s Review Service, August, 2010)

From the Publisher