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TitleAdult Stem Cells - Biology and Methods of Analysis - D. Phinney (Humana, 2011) WW
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Total Pages290
Table of Contents
Adult Stem Cells
ISBN 9781617790010
Part I:
Basic Biology of Adult Stem Cells
	Chapter 1: Molecular Mechanisms Regulating Adult Stem Cell Self-Renewal
		1.1 Tissue Homeostasis and the Stem Cell Concept
		1.2 Methods for Analysis of Stem Cells and Derivatives
			1.2.1 Clonal Analysis
			1.2.2 Prospective Isolation
			1.2.3 Additional Techniques
		1.3 Determining Location, Origin, and Function of Adult Stem Cells: The Brain as an Example
			1.3.1 Location and Identity
			1.3.2 Spatial/Temporal Origins and Clonal Diversity
			1.3.3 Function
		1.4 Cellular Properties and Determinantsof Self-Renewal and Potency
			1.4.1 Mitogens and Morphogens
			1.4.2 Lateral Inhibition and Stem Versus Not-Stem Cell Fate Choice
			1.4.3 Coordination of Cell Cycle, Potency and Self-Renewal
			1.4.4 Transcription Factors Regulating Self-Renewal in Development and Throughout the Lifespan
			1.4.5 Adult Functions of Self-Renewal Factors and the Balance of Quiescence and Exhaustion
		1.5 Summary and Perspectives
	Chapter 2: Maintenance of Adult Stem Cells: Role of the Stem Cell Niche
		2.1 Introduction
		2.2 Quiescence and Self-Renewal
		2.3 The Hematopoietic Stem Cell Niche
			2.3.1 Niche Cell Components Two Types of Niches Close Interaction Between the Endosteal and Perivascular Zones
			2.3.2 Niche Factors Adhesion Molecules Cytokines and Growth Factors Local Environmental Factors
		2.4 Intrinsic Programs in HSCs
		2.5 Mobilization of Normal and Leukemic Stem Cells
		2.6 Concluding Remarks
	Chapter 3: The Emerging Role of microRNAs in Adult Stem Cells
		3.1 Introduction
		3.2 microRNA Biogenesis
		3.3 microRNA Mechanisms of Action
		3.4 The Role of miRNAs in Embryonic Stem Cells
		3.5 The Role of miRNAs in Differentiation of Adult Stem Cells
			3.5.1 ES to EB Transition: The Beginnings of ASCs
			3.5.2 Cardio- and Myogenesis
			3.5.3 Neurogenesis
			3.5.4 Hematopoiesis
			3.5.5 Osteogenesis
			3.5.6 Epitheliopoiesis
			3.5.7 Germ Cells
			3.5.8 Loss of Self-Renewal
		3.6 miRNAs in Cellular Stress Disease, and Trauma
			3.6.1 Cellular Stress and Apoptosis
			3.6.2 Cancer
			3.6.3 Spinal Cord Injury
			3.6.4 Ethanol Exposure in the Fetal Brain
			3.6.5 Polyglutamine-Induced Neurogenesis
			3.6.6 Autoimmunity
		3.7 Miscellaneous miRNA-Related Disorders
		3.8 Brief Description of miRNA Protocols
		3.9 Conclusions
	Chapter 4: Expression and Function of Pluripotency Genes in Adult Stem Cells*
		4.1 POU5F1 (Also Known as OCT3, OCT4, OTF3 or OTF4)
			4.1.1 Introduction
			4.1.2 Oct4 Pseudogenes
			4.1.3 Oct4 Gene and Isoforms
			4.1.4 Oct4 in Nonembryonic Stem Cells Testis-Derived Stem Cells Somatic Tissue-Derived Stem Cells
			4.1.5 Oct4 in Somatic Tumors and Transformed Cell Lines
		4.2 NANOG
			4.2.1 Introduction
			4.2.2 Nanog Pseudogenes and Alternative Splice Forms
			4.2.3 Nanog Expression in Adult Cells
		4.3 Conclusion
	Chapter 5: Adult Stem Cell Plasticity Revisited
		5.1 Introduction
		5.2 Is There Transdifferentiation?
		5.3 Contribution of Bone Marrow-Derived Stem Cells to Cells of Other Organs
			5.3.1 Bone Marrow Differentiation into Organs of Ectodermal Origin CNS Skin/Hair Follicles/Fingernails
			5.3.2 BM Differentiation into Organs of Endodermal Origin Oral Mucosa Uterine Endometrium Gastrointestinal (GI) Tract Liver Pancreas Lung Kidney
			5.3.3 BM Differentiation into Organs of Mesodermal Origin Skeletal Muscle Cardiac Muscle
		5.4 Summary
Part II:
Characterization of Adult Stem Cells
	Chapter 6: Lineage Tracing of Tissue-Specific Stem Cells In Vivo
		6.1 Incorporation of DNA Nucleoside Analogs and Chromatin Manipulation
			6.1.1 Tritiated Thymidine
			6.1.2 Chlorodeoxyuridine, Bromodeoxyuridine, and Iododeoxyuridine
			6.1.3 Ethynyl Deoxyuridine
			6.1.4 Tagged Histone Proteins
		6.2 Cellular Marking Through Genetic Reporter Strategies
			6.2.1 Mosaics Generated Through Retroviral Tagging
			6.2.2 Mosaicism Through Cre Recombinase-Mediated Site-Specific Recombination Transgenic Systems for Drug-Regulated Expression of Cre Ligand-Regulation of Cre Activity Limitations of Cre-lox Systems
			6.2.3 Light-Emitting Reporters Green Fluorescent Protein Fluorescent Protein Variants Combined Fluorescent Protein Expression (Brainbow Mice) Luciferase
			6.2.4 Chimera Models Hematopoietic Stem Cell Origin of Chimeric Models Embryonic Models of Chimerism Chimerism Via Delivery of Hematopoietic Stem Cells to Adult Organisms Lineage Tracing in Human Mitochondrial DNA
			6.2.5 Phylogenetic Fate Mapping
		6.3 Ectopic Stem Cell Implantation
			6.3.1 Renal Capsular Assays
			6.3.2 Implantation Models Using Biomatrices
			6.3.3 Improving Ectopic Implantation Methods
	Chapter 7: Surrogate Measures of Adult Stem Cell Self-Renewal: The Neural Stem Cell Paradigm
		7.1 Introduction
		7.2 Measuring NSC Activity In Vitro
			7.2.1 The Neurosphere Assay, a Gold Standard Methodology
			7.2.2 Limitations of the Neurosphere Assay
			7.2.3 The Neural Colony-Forming Cell Assay to Enumerate Actual Neural Stem Cells
		7.3 Stem Cell Niche Repopulation In Vivo
		7.4 “Quantum” Stem Cell Mechanics
		7.5 Summary
	Chapter 8: ABC Transporters, Aldehyde Dehydrogenase, and Adult Stem Cells
		8.1 Introduction
		8.2 Aldehyde Dehydrogenases
			8.2.1 ALDH Activity in Haematopoietic Cells
			8.2.2 ALDH Activity in Non-Haematopoietic Organs
		8.3 The Side Population (SP)
			8.3.1 SP Cells in Haematopoietic Cells
			8.3.2 SP Cells in Skeletomuscular Tissues
			8.3.3 SP Cells in Epithelia
		8.4 Conclusions
Part III:
Regulation of Life Spanand Immortalization
	Chapter 9: Regulation of Life Span in Adult Stem Cells
		9.1 Introduction
			9.1.1 Aging and Life Span of Somatic Cells
			9.1.2 Aging and Life Span of Stem Cells
			9.1.3 Telomeres and Cell Senescence
		9.2 Intrinsic Mechanisms of Life Span Regulation in Stem Cells
			9.2.1 Accumulation of DNA Damage
			9.2.2 Reactive Oxygen Species
			9.2.3 Epigenetic Alterations
			9.2.4 Telomeres and Telomerase
		9.3 Extrinsic Mechanisms of Life Span Regulation in Stem Cells
		9.4 Mechanisms of Termination of Stem Cell Life Span
		9.5 Therapeutic Approaches to the Regulation of Stem Cell Life Span
			9.5.1 Methods to Extend Stem Cell Life Span
			9.5.2 Methods to Shorten Stem Cell Life Span
	Chapter 10: The Cancer Stem Cell Paradigm
		10.1 Introduction
		10.2 History of the Cancer Stem Cell Hypothesis
		10.3 Stem-Like Behaviors and Signaling Provide Selective Advantages for Cancer Cells and Tumor Growth
		10.4 Cancer Stem Cell Properties
		10.5 Isolation and Identification of Cancer Stem Cells
		10.6 In Vitro and In Vivo CSC Maintenance
		10.7 Debate Surrounding Cancer Stem Cells
			10.7.1 Terminology
			10.7.2 Rarity
			10.7.3 Cellular Origin of Tumors
			10.7.4 What Makes a CSC? Genomic Alteration vs. Epigenomic Alteration vs. Plasticity
		10.8 Glioblastoma: Deadly Tumor and Model Systemfor Solid Tumor Cancer Stem Cells
		10.9 Conclusions and Implications for Therapeutic Approaches
Name Index
Subject Index
Document Text Contents
Page 2

Stem Cell Biology and Regenerative Medicine

Series Editor
Kursad Turksen, Ph.D.
[email protected]

For other titles published in this series, go to

Page 145


Page 146

135D.G. Phinney (ed.), Adult Stem Cells: Biology and Methods of Analysis,
Stem Cell Biology and Regenerative Medicine, DOI 10.1007/978-1-61779-002-7_6,
© Springer Science+Business Media, LLC 2011

Abstract Tissue-specific stem cells are characterized by their ability to undergo
unlimited self-renewal and generate transit-amplifying progeny that yields all spe-
cialized cell types of a tissue. Both identification of stem cells and characterization
of their properties have been possible through the use of a range of methods to track
cell fate in vivo or in vitro. In vivo systems for lineage tracing offer the advan-
tage of keeping stem cells within their local microenvironment affording them
exposure to signaling molecules that help govern their tissue-specific behavior;
this will serve as the focus of discussion within this chapter. Multiple methods of
lineage tracing in vivo will be described. These methods account for difficulties
of lineage tracing within complex organ systems such as analysis of rare numbers of
cells, large variability in the type and number of differentiated cell types, impaired
visualization of cell type-specific markers in solid organs, and differences between
organs in the rate of cell turnover. Methods discussed will include classical methods
such as the incorporation of DNA nucleoside analogs or chromatin manipulation.
This will be followed by discussion of more contemporary methods focused on
the incorporation of genetic reporters through mosaic or chimera models and the
implantation of ectopic stem cells. Although these approaches are described
individually in the following sections, they are often applied in combination to
overcome specific limitations of individual methods and to more rigorously define
stem cell behavior in vivo.

Keywords Adult stem cells • Asymmetric cell division • Cre-lox • DNA replication
• DNA nucleoside analogs • Green fluorescent protein • Label retention • Light-emitting
reporters • Lineage tracing • Self-renewal

B.R. Stripp (*)
Department of Medicine, Division of Pulmonary and Critical Care Medicine,
Duke University Medical Center, Durham, NC 27710, USA
Department of Cell Biology, Division of Pulmonary and Critical Care Medicine,
Duke University Medical Center, Box 103000 DUMC, Durham, NC 27710, USA
e-mail: [email protected]

Chapter 6
Lineage Tracing of Tissue-Specific
Stem Cells In Vivo

Kurtis T. Sobush, Keitaro Matsumoto, Huaiyong Chen, and Barry R. Stripp

Page 289

278 Subject Index

Renal capsular assays, 155–156
Repressor element 1 (RE1) silencing

transcription factor (REST), 64–65
Retroviral vector labeling, 141–142
Reverse-tet-transactivator (rtTA), 145–146
RNaseIII enzyme, 59–60

Side population (SP)

analysis, pancreatic cancer cell lines, 189
definition, 188

endocrine tissues, 192
endometrial, 192
epithelial cells, 193
eye, 194
mouse and human kidney, 191–192
undifferentiated, 193

features, 188–189
haematopoietic cells, 190
phenotype, 188
skeletomuscular tissues

rodent heart, 191
satellite SP, 190–191
soft tissues, 191

of solid organs, 189
Signaling lymphocyte activation molecule

(SLAM), 41
Skeletomuscular tissues

rodent heart, 191
satellite SP, 190–191
soft tissues, 191

Sonic Hedgehog (Shh), 17–18
Spermatogonial progenitor cells (SPCs), 99
Spermatogonial stem cells (SSCs), 99–100
Spinal cord injury (SCI), 81–82
Stem and progenitor cells, functional

properties, 170, 171
Stem cell niche

fate regulation, 38
functions, 37

components, BM, 44
endosteal vs. perivascular zones, 43–44
factors, 44–48
intrinsic programs, 48–49
mobilization and leukemic stem cells,

mobilization schema, 50, 51
quiescent, 37
self-renewal, 38–39
structure models, 40, 42
types, 40–43

regulation, importance, 51
types, stem cell, 37

Stem cells
adult, developmental origin, 11

advantages, 27–28
5-bromo–2-deoxyuridine (BrdU)

labelling, 7
clonal, 5–6
conditional/inducible gene targeting, 7
immunocytochemistry, 6–7
prospective isolation, 6
in vitro and in vivo, 5

cellular properties and determinants
cell cycle, 22–24
determination, 16
lateral inhibition, 19–20
mitogens, 16–17
morphogens, 17–19
notch signaling, 20–22
selection, 16
self-renewal capacity, 14–16
self-renewal factors, adult functions,

stem vs. not-stem cell fate, 19–22
transcription factors, 24–25

characteristics, 36
clonal diversity, 12–14
discovery, 36
function, 14
location and identity

adult neurogenesis, 8
progenitor cells, 10
quiescence, 10
type B cells, 8–10

reconstitution, 4
selective vs. instructive actions, 16
spatial/temporal origins, 10–14
stemness, 27

Stochastic theory, tumor evolution, 226
Strontium (Sr), 42
Subventricular zone (SVZ), 8, 10–14,

Symmetric division, stem cell, 15

Telomerase, 206–208
Tet operon, 143–146
Tet-transactivator (tTA), 145
Thrombopoietin (THPO), 46–47
Tissue homeostasis, 4–5
Tissue-specific stem cells, in vivo lineage


Page 290

279Subject Index

cellular marking, genetic reporter
chimera models, 151–154
Cre recombinase/Cre-lox site-specific

recombination, 142–148
light-emitting reporters, 148–151
methods, 141
phylogenetic fate mapping, 154–155
retroviral vector labeling, mosaic

generation, 141–142
DNA nucleoside analogues and chromatin

CldU, BrdU and IdU, 138–139
ethynyl deoxyuridine (EdU), 139–140
tagged histone proteins, 140
tritiated thymidine (H3-thymidine), 137

ectopic stem cell implantation
development, 155
Matrigel, 156
renal capsular assays, 155–156
research on, 156

Totipotency, 15
Tritiated thymidine (H3-thymidine), 137
Tumor necrosis factor-a (TNFa), 45

Uncertainty principle, “Quantum” stem cell

mechanics, 172

Vascular endothelial growth factor

(VEGF), 240
Ventricular zone (VZ), 12–13
Very small embryonic like (VSEL) cells,


Wingless/Int proteins, 18

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