Isolation, Characterization and Utilization of CNS Stem Cells

202 Pages·1997·5.576 MB·English

by  I. L. Weissman (auth.), F. H. Gage Ph. D., Y. Christen Ph. D. (eds.)

RESEARCH AND PERSPECTIVES IN NEUROSCIENCES
Fondation Ipsen
Editor
Yves Christen, Fondation Ipsen, Paris (France).
Editorial Board
Albert Aguayo, McGill University, Montreal (Canada).
Philippe Ascher, Ecole Normale Superieure, Paris (France).
Alain Berthoz, College de France, CNRS UPR 2, Paris (France).
Jean-Marie Besson, INSERM U 161, Paris (France).
Emilio Bizzi, Massachusetts Institute of Technology, Boston (USA).
Anders Bjorklund, University of Lund, Lund (Sweden).
Ira Black, University of Medicine & Dentristry of New Jersey, Piscataway (USA).
Floyd Bloom, Scripps Clinic and Research Foundation, La Jolla (USA).
Joel Bockaert, Centre CNRS-INSERM de Pharmacologie Endocrinologie,
Montpellier (France).
Pierre Buser, Institute des Neurosciences, Paris (France).
Jean-Pierre Changeux, College de France, Institut Pasteur, Paris (France).
Carl Cotman, University of California, Irvine (USA).
Stephen Dunnett, University of Cambridge, Cambridge (UK).
George Fink, Medical Research Council, Edinburgh (UK).
Fred Gage, Salk Institute, La Jolla (USA).
Jacques Glowinski, College de France, Paris (France).
Claude Kordon, INSERUM U 159, Paris (France).
Michel Lacour, CNRS URA 372, Marseille (France).
Michel Le Moal, INSERM U 259, Bordeaux (France).
Gary Lynch, University of California, Irvine (USA).
Brenda Milner, McGill University, Montreal (Canada).
John Olney, Washington University Medical School, Saint Louis (USA).
Alain Privat, INSERM U 336, Montpellier (France).
Allen Roses, Duke University Medical Center, Durham (USA).
Constantino Sotelo, INSERM U 106, Paris (France).
Jean-Didier Vincent, Institute Alfred Fessard, CNRS, Gif-sur-Yvette (France).
Bruno Will, Centre de Neurochimie du CNRS/INSERM U 44 Strasbourg (France).
Springer
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F. H. Gage Y. Christen (Eds.)
Isolation, Characterization
and Utilization
of CNS Stem Cells
With 43 Figures, Some in Color and 5 Tables
Springer
Gage, F. H., Ph. D.
Laboratory of Genetics
The Salk Institute for Biological Studies
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San Diego, CA 92186-5800
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Christen, Y., Ph. D.
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ISBN-13: 978-3-642-80310-9 e-ISBN-13: 978-3-642-80308-6
001: 10.1007/978-3-642-80308-6
Library of Congress Cataloging-in-Publication Data. Isolation, characterization, and utilization of CNS
stem cells/F. H. Gage, Y. Christen (eds.). p. cm. - (Research and perspectives in neurosciences) Includes
bibliographical references and index. ISBN 3-540-61696-9 (alk. paper) 1. Developmental neurophy
siology, 2. Stem cells. 3. Central nervous system - Growth. I. Gage, F. (Fred), 1950-II. Christen, Yves.
III. Series. QP356.25.I86 1996 612.8'2-dc20 96-38498 (CIP)
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Contents
Stem Cells: The Lessons from Hematopoiesis
1.L. Weissman ..................................................... .
NRSF: A Coordinate Repressor of Neuron-Specific Genes Expressed in CNS
Neural Progenitor Cells
C.J. Schoenherr and D.J. Anderson ...... . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9
Segregation of Cell Lineage in the Avian Neural Crest
E.Dupin, C. Ziller, and N.M. Le Douarin . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 29
Comparative Strategies of Sub ependymal Neurogenesis in the Adult Forebrain
S.A. Goldman. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 43
Characterization of Neuronal Progenitor Cells of the Neonatal Forebrain
M.B. Luskin, T. Zigova, R. Betarbet, and B.J. Soteres . . . . . . . . . . . . . . . . . . . .. 67
Neurogenesis in the Adult Brain: Prospects for Brain Repair
A. Alvarez-Buylla... . ... . . . ........ . . .... . . . . .. .... . ..... ....... . . .. 87
The Oligodendrocyte-Type-2 Astrocyte Lineage: In vitro and in vivo Studies
on Development, Tissue Repair and Neoplasia
M. Noble .......................................................... 101
Neurogenesis in the Adult Brain: Lessons Learned from the Studies of
Progenitor Cells from the Embryonic and Adult Central Nervous Systems
J. Ray, T.D. Palmer, J. Suhonen, J. Takahashi, and P.R. Gage. . . . . . . . . . . . . .. 129
Use of Conditionally Immortalized Neural Progenitors for Transplantation
and Gene Transfer to the CNS
A. Martinez-Serrano, C. Lundberg, and A. Bjorklund .................... 151
The Origins of the Central Nervous System
R. McKay ......................................................... 169
Transplantation and Differentiation of Neural "Stem-Like" Cells:
Possible Insights into Development and Therapeutic Potential
E. Y. Snyder, J.D. Flax, B.D. Yandava, K.1. Park, S. Liu, C.M. Rosario,
and S. Aurora ...................................................... 173
Subject Index ...................................................... 197
Preface
This volume is based on a meeting of the Fondation IPSEN, held in Paris on Sep
tember 18, 1995 to address the main issues of nervous system stem cells biology.
Cell replacement in the adult mammals is not unusual outside the nervous sys
tem. In fact, the nervous system is unique in lacking the ability to replace cells,
following damage. Most neurons, in the adult central nervous system are termin
ally differentiated, exist through the life of the organism and are not replaced
when they die. There are, however, regions of the postnatal brain that continue to
produce new neurons, but the fate and longevity of those cells are not well
known. Evidence exists that small populations of neurons continue to be born in
the adult ventricular zone, olfactory epithelium and hippocampus. In the adult
hippocampus, newly born neurons originate from putative stem cells that exist in
the sub granular zone of the dentate gyrus. Progeny of these putative stem cells
differentiate into neurons in the granular layer within a month of the cells' birth,
and this late neurogenesis continues throughout the adult life of the rodent.
By understanding the nature of progenitor cells present in the embryonic and
adult brains, the change in their population dynamics during development, and
the factors that influence their proliferation, fate choice and differentiation, it
may be possible to develop a strategy to manipulate cells in situ to treat neuro
degenerative diseases or the injured adult brain.
Fred GAGE
Yves CHRISTEN
Aknowledgments: The editors wish to thank Mary Lynn Gage for editorial assist
ance and Jacqueline Mervaillie for the organization of the meeting in Paris.
Stem Cells: The Lessons from Hematopoieses
I.L. Weissman
The driving force that led to the isolation of hematopoietic stem cells (HSCs) was
primarily medical, and not simply biological, inquiry. Following the massive
exposure of the civilian populations to ionizing radiation in 1945, experimental
animal models of whole body lethal radiation soon revealed that the deaths that
occur at the lowest lethal doses were the result of a disrupted hematopoietic sys
tem: the loss of granulocytes led to infection, the loss of platelets led to bleeding,
and the loss of red blood cells led to fatal anemias. This fatal radiation syndrome
could be prevented by shielding a single long bone, or by injecting bone marrow
from identical twin mice into the irradiated host. Two sets of discoveries set the
stage for the isolation of HSCs: 1) In 1956 three groups demonstrated that bone
marrow (chromosomally marked) injected into lethally irradiated hosts saved the
hosts by reconstituting the host hematolymphoid system with donor-derived
cells (Ford et al. 1956; Makinodan 1956; Nowell et al. 1956). 2) In 1961 while
seeking an assay to understand the potential different effects of x-rays on normal
versus neoplastic cells, Till and McCulloch (1961) observed that limiting doses of
bone marrow injected into lethally irradiated mice resulted in the appearance of
spleen colonies, each colony containing several different cell types in the myelo
erythroid series. Pre-irradiation of the donor resulted in random chromosomal
translocations in donor bone marrow (BM); limiting doses of that marked BM
resulted in a spleen colony (CFU-S) within which all cells bear the same unique
translocation. Till and McCulloch concluded that the diverse cell types contained
within the single spleen colony derive from a single clonogenic precursor (Till
and McCulloch 1963; Wu et al. 1967). The cells in a single spleen colony could be
retransferred to a secondary host to form, again, multilineage spleen colonies
(Wu et al. 1967; Siminovitch et al. 1963) as well as donor-derived lymphoid cells
(Lafleur et al. 1972; Wu et al. 1968); these collective findings led McCulloch and
Till to propose the hypothesis that there are a small number of HSCs in a bone
marrow transplant that can lead to reconstitution of hematopoiesis, and, at the
single cell level, these HSCs can self renew as well as give rise to multilineage dif
ferentiation (Till and McCulloch 1963). Thus two major issues in the practice of
medicine - radioprotection from whole body lethal irradiation, and the use of
radiotherapy to treat local tumors - set the stage for the characterization and
isolation of HSCs. I believe that both rational scientific inquiry and medical need
will drive the isolation of stem cells from other organ systems.
F. Gage I Y. Christen (Eds.)
Isolation, Characterization and Utilization of CNS Stem Cells
© Springer-Verlag Berlin Heidelberg 1997
2 1.1. Weissman
Why were HSCs the first stem cell type to be isolated from any tissue? I
believe that this resulted not only from the medical issues described above, but
also because cells in the bone marrow reside as loosely associating cell types,
easily brought into suspension. They give rise to other cell types easily identified
by morphology or phenotype, also largely unconnected to other cells, and there
fore analyzable in cell suspensions. Also, hematopoietic cells, including HSCs,
when injected back into the blood stream home to the irradiated and appropriate
microenvironment with high efficiency (Weissman et al. 1978) alllowing a reduc
tionist, quantitative analysis of clonogenic and radioprotective cells in this sys
tem not clearly available in other systems. But why do HSCs placed into the blood
of an irradiated mouse home so efficiently to bone marrow and splenic hemato
poietic microenvironments? The simplest answer is that it is probably a behavior
that occurs frequently, and therefore has some survival value. Hematopoiesis in
vertebrates occurs mainly in the bone marrow and also, in some species, in the
spleen. In embryonic life, hematopoiesis is established in the yolk sac and near
the dorsal aorta or the gonad and mesonephros, moving to the fetal liver during
fetal life, and then to the bone marrow (and in mouse the spleen, as well) in late
fetal and postnatal life (Muller et al. 1994). We have shown that cells in early
embryonic yolk sac blood islands can establish the adult, bone marrow-contered
hematolymphoid system (Weissman et al. 1977, 1978). We have also shown that
the secondary movement of HSCs out of the murine fetal liver occurs at a time
when they appear in fetal spleen and bone marrow (Morrison et al. 1995). Thus
is appears that there are spontaneous movements of HSCs from one organ to
another, necessitating not only a passage through the blood but also a de
adhesion from the initial site and a homing to the secondary site. Each of these
events is almost certainly controlled by surface adhesion molecules on stem cells
and their counter ligands in the hematopoietic stroma of these micro environ
ments, as well as the blood vessels that line them. In this view, HSC adhesion
molecules first recognize vascular addressins and then counter receptors (e.g.,
VCAM-1) on hematopoietic stroma (Papayannopoulou et al. 1995; Miyake et al.
1991; Kina et al. 1991). Remarkably, these events occur time and again in adult
life at a very low level as the hematopoietic stem cells and committed progeny
leave the bone marrow to enter the blood and home to the thymus, spleen, and
other bone marrow sites (Hanks 1963, Lepault and Weissman 1981). Mobilization
is greatly augmented when host hematopoiesis is largely destroyed by sublethal
doses ofx-ray, chemotherapeutic agents such as cyclophosphamide or the type of
endotoxic shock that follows several types of infectious episodes (Molineux et al.
1990; Vos et al. 1972; Gianni et al. 1989). The loss of hematopoietic cells presum
ably results in the induced expression of hematopoietic cytokines, which along
with the cytotoxic agent leads to the mobilization of surviving HSCs and other
progenitors from the bone marrow into the blood, followed by the seeding of
other intact hematopoietic sites (Molineux et al. 1990, To et al. 1984). In the
mouse the increase in mobilized HSCs in secondary organs such as the spleen
(D Wright, SJ Morrison, IL Weissman, unpublished observations) presumbly occurs
by rehoming of mobilized cells, but could also involve self-renewing divisions by
Stem Cells: The Lessons from Hematopoieses 3
HSCs. Because external radiation, although uncommon, can occur in the natural
environment, and more commonly toxic compounds capable of killing dividing
cells are produced by fungi, plants, and other organisms that can be ingested, it
is not surprising that HSCs can be mobilized for the purposes of re-establishing
stem cell niches, hematopoiesis, and lymphopoiesis.
What are the lessons that can be applied when studying the dynamics and
characteristics of stem cells from other organs? The obvious issues are to find
those cases of tissue formation or organogenesis in which progenitor stem cell
migration occurs, and to understand the role of progenitors in the processes of
repair and regeneration of damaged organs. For example, germ cells also
undergo natural migrations during development, appearing in the yolk sac dur
ing embryonic life, and then migrating to the genital ridges during fetal life
(Mintz 1957). Neural crest cells also follow extensive pathways of migration of
form body components as diverse as smooth muscle cells, adrenal medullary
cells, melanocytes, and cells of the peripheral nervous system (Anderson 1993;
Le Douarin et al. 1993). Strikingly, primitive hematopoietic cells, germ cells, and
neural crest cells all share the cell surface signaling transmembrane tyrosine
kinase called c-kit, and stromal cells that interact with them produce cell surface
and soluble ligands for the kit receptor, SLF (Huang et al. 1992). Thus c-kit can
also be used as an identifier of primitive cells in these three organ systems,
although it is not expressed solely by stem/progenitor cells. We (Ikuta and Weiss
man 1992) and others (Ogawa et al. 1991; Orlic et al. 1993) have shown that
hematopoietic stem cells as well as early hematopoietic progenitors are c-kit+.
Techniques similar to those used for the identification and isolation of hemato
poietic stem cells have been used to isolate neural crest stem cells (Stemple and
Anderson 1992), and presumably such techniques should be valuable in isolating
migrating stem cells of the germ line as well. However, in most epithelial (and
perhaps neuroepithelial) organs and tissues, cells are held together by tight junc
tions. Although it is not clear that these tight cell-cell adhesions are shared by the
stem cells of these organs, it appears clear for most of the these systems that stem
cells are present within the defined organ and do not migrate in from other sites.
Thus, stem cells might be harvested during conditions that activate and mobilize
stem cells from these organ systems, such as fetal development and regeneration.
The Isolation of HSCs
The discovery of spleen colonies (see above) formed the intellectual basis for
assays leading to the identification and purification of HSCs. Because HSCs must
include clonal progenitors of all cell lineages, and also self-renew, the establish
ment of clonogenic assays for cells other than myeloerythroid cells was the first
step. In the mid 1970s, T. Michael Dexter and colleagues (1977) found that bone
marrow stroma could support hematopoiesis in vitro. In the early 1980s, With
lock and Witte (1982) modified the Dexter culture system in a way that permitted
lymphopoiesis of B lymphocytes. Following her work with Witte, Cheryl Witlock
4 I.L. Weissman
joined my laboratory. We cloned the stromal cells necessary for establishment of
B lymphopoiesis and showed that bone marrow cells added at limit dilution to
these defined stromal lines could form colonies (Whitlock et al. 1987). The subset
of bone marrow cells that were clonogenic lacked surface markers of all known
mature blood cell types (Muller-Sieburg et al. 1986). Starting with a cocktail of
antibodies that identified mature blood cell surface markers, we used negative
and positive selection techniques to identify the cell surface phenotype of the
Whitlock/Witte culture initiating cell (Muller-Sieburg et al. 1986). Comparing the
clonogenic cells for the Whitlock/Witte stromal culture system and the Tilll
McCulloch CFU-S, we found that there, was a subset of cells that shared this phe
notype (Muller-Sieburg et al. 1986). Over the same interval we had been studying
thymic lymphopoiesis of T cells and had developed a clonal assay for thymic pro
genitors (Lepault and Weissman 1981; Ezine et al. 1984; Spangrude and Weiss
man 1988; Rouse and Weissman 1981). We were able to show that a rare popula
tion (0.05 % of bone marrow or 1 in 2000 cells) contained clonogenic progenitors
for all three assays, covering all known mature blood cell types (Spangrude et al.
1988). In mice the constellation of markers that identify hematopoietic stem cells
are Thy 1.11°, Lin-!1o, Sca1(LY6A/E} and c-kit+ (Ikuta and Weissman 1992; Spang
rude et al. 1988). Remarkably, these cells represent 1 in 2000 cells in bone marrow
and are I-to 2000-fold enriched for radioprotective cells (Ikuta and Weissman
1992; Spangrude et al. 1988); they give rise to both short-term and long-term
hematopoiesis that is donor derived for all lineages (Uchida et al. 1994), as well
as renewing up to 150,000 HSC progeny (Spangrude et al. 1991; Smith et al. 1991;
Morrison and Weissman 1994). One to 20 genetically marked HSCs can be mixed
in with a 1000 radioprotective host strain HSCs and injected back into lethally
irradiated hosts to determine the full developmental potential of a single stem
cell (Smith et al. 1991; Morrison and Weissman 1994). These studies have
revealed not only that single HSCs are multilineage progenitors, but also that a
subset of them has high self-renewal capacity and prolonged productivity,
whereas the majority of HSCs have multilineage differentiation capacity but
more limited self renewal, and therefore a shorter life span (Morrison and Weiss
man 1994). The long-term HSCs (LT-HSCs) can be differentiated from short
term HSCs and multipotent progenitors without stem cell self-renewal capacity
not only in terms of their life span but also in terms of their surface makers (Mor
rison and Weissman 1994). Thus long-term and short-term HSCs can be isolated
in mouse and now in man (Baum et al. 1992) and their developmental potential
(Tsukamoto et al. 1995), the specific genes that they express in one versus
another state of development, their niche relationships, and their role in bone
marrow transplantation recovery of essential cell elements at early and late inter
vals can be studied. One of the more intriguing differences between stem cells
with a long-term, productive life span and those with a short productive life span
is in the gene complex that controls rebuilding telomeres in dividing cells
(SJ Morrison, S Prowse, P Howe, IL Weissman, in press, Immunity). The RNA
priming of DNA replication theoretically leaves the 5' end of telomeres with a gap
representing the length of the RNA primer, but in many dividing cells this gap is