Tài liệu Embryonic stem cells recent advances in pluripotent stem cellbased regenerative medicine - Edited b

EMBRYONIC STEM CELLS RECENT ADVANCES IN PLURIPOTENT STEM CELLBASED REGENERATIVE MEDICINE
Edited by Craig S. Atwood




Contents
Preface IX
Part 1 Clinical Applications and Ethical Considerations 1
Chapter 1 The Role of Complicity in the Ethics of Embryonic Stem Cell Research 3
Dieter Birnbacher
Chapter 2 Potential Clinical Applications of Embryonic Stem Cells 21
Arianna Malgieri, Giuseppe Novelli and Federica Sangiuolo


Part 2 Tissue-specific Regeneration of the Heart 49
Chapter 3 Chemical Biology of Pluripotent Stem Cells: Focus on Cardiomyogenesis 51
Jijun Hao, Li Zhou and Charles C. Hong
Chapter 4 Cardiac Differentiation of Embryonic Stem Cells by Patterning Culture 65
Daisuke Sasaki and Teruo Okano


Part 3 Tissue-specific Regeneration of the Brain and Sensory Organs 81
Chapter 5 Embryonic Stem Cell-Derived Multipotent Mesenchymal Stromal Cell Therapy Following Focal Ischemia in the Rat 83
Gregory C. Kujoth and Mustafa K. Başkaya

Chapter 6 Embryonic Stem Cell in the Therapy of Neurodegenerative Diseases 105
Xiaotang Fan, Yongping Tang, Kai Wang, Xiang Cui, Shiqi Tao and Haiwei Xu
Chapter 7 Engineering Therapeutic Neural Stem Cell Lines for Parkinson’s Disease 137
Marcel M. Daadi
Chapter 8 Embryonic Stem Cells Overexpressing the Recognition Molecules L1 and Tenascin-R Enhance Regeneration in Mouse Models of Acute and Chronic Neurological Disorders 149
Gunnar Hargus and Christian Bernreuther
Chapter 9 Perspectives of Stem Cell-Derived Microglia for Medicine 171
Kristin Roy, Clara Beutner and Harald Neumann
Chapter 10 Embryonic Stem Cell-Derived Neurons for Inner Ear Therapy 189
Eri Hashino and Michael H Fritsch
Chapter 11 Potential of Pluripotent Stem Cells for the Replacement of Inner Ears 203
Koji Nishimura, Takayuki Nakagawa and Juichi Ito
Chapter 12 Stem Cells and the Retina – Challenges for Regenerative Medicine 211
Andrea Messina, Simona Casarosa and Elisa Murenu


Part 4 Tissue-specific Regeneration of Hematopoietic Systems 237
Chapter 13 Generation of Blood Cells from Human Embryonic Stem Cells and Their Possible Clinical Utilization 239
Feng MA, Wenyu Yang, Yasuhiro Ebihara and Kohichiro Tsuji
Chapter 14 Hematopoietic Differentiation from Embryonic Stem Cells 251
Yasuhisa Yokoyama, Hidekazu Nishikii and Shigeru Chiba
Chapter 15 ES Cell-derived Erythroid Cell Lines Able to Produce Mature Red Blood Cells 273
Yukio Nakamura


Part 5 Tissue-specific Regeneration of Other Tissues 289
Chapter 16 Differentiation of Hepatocytes from Mice Embryonic Stem Cells in Three-Dimensional Culture System Imitating in vivo Environment 291
Tetsuya Imamura
Chapter 17 Rejuvenation of the Thymic Microenvironment by ESC-derived Thymic Epithelial Progenitors 301Laijun Lai

Chapter 18 Actual Achievements on Germ Cells and Gametes Derived from Pluripotent Stem Cells 311
Irina Kerkis, Camilla M. Mendes, Simone A. S. da Fonseca, Nelson F. Lizier, Rui C. Serafim and Alexandre Kerkis
Part 6 Side Effects of Pluripotent Stem Cell Therapies 337
Chapter 19 Self-Renewal, Pluripotency and Tumorigenesis in Pluripotent Stem Cells Revisited 339
Yanzhen Li and Tetsuya S. Tanaka
Chapter 20 Generation of Human Induced Pluripotent Stem (iPS) Cells from Liver Progenitor Cells by Two Chemicals and the Clinical Application 359
Hisashi Moriguchi, Makoto Mihara, Chfumi Sato and Raymond T Chung

Chapter 21 Embryonic and Cancer Stem Cells - two views of the same landscape 371
Unai Silván, Alejandro Díez-Torre, Lucía Jiménez-Rojo and Juan Aréchaga
Chapter 22 Genome Stability in Embryonic Stem Cells 399
Paola Rebuzzini, Maurizio Zuccotti, Carlo Alberto Redi and Silvia Garagna






Preface
The hope that one day human pluripotent stem cells (hPSCs) can be utilized for the
treatment of various diseases has swept throughout the world since the isolation and
in vitro maintenance of human embryonic stem cells (hESCs) just before the end of
the last millennium (Thomson et al. 1998). Contrasting with this tremendous hope of
hESCs for regenerative medicine and continued mortality lies the moral issues related
to the use of pluripotent ESCs obtained from the discarded embryos of in vitro fertilization
(the termination of life). In the fi rst section of this book, ‘Clinical Applications
and Ethical Considerations’ the fi rst chapter by Malgieri et al. highlights the potential
for stem cell technologies, including hESC technologies, in regenerative medicine. The
second chapter by Birnbacher delves into the complex bioethical and biopolitical issues
of using hESCs for basic and applied applications. Birnbacher provides a thoughtful
and in depth assessment of complicity- the role of scientists as accomplices in the
propagation of a “wrong doing”. As Birnbacher points out, the wrong doing is not so
much the use of ESCs, themselves not human embryos, “but moral concerns as to the
source from which the stem cells are derived and the methods by which they are retrieved
(or the life terminated).” As the authors continue, “Complicity is the core of the
criticism levelled against research on pluripotent hESC in many quarters and has even,
in some countries, become the basis of legal prohibitions. Complicity is a problem only
for those who are torn between the conviction that embryo research is (for intrinsic or
extrinsic reasons) a moral evil and the conviction that hESC research is worth pursuing
either for its medical or for its scientifi c prospects or both. The class most likely to
face this uncomfortable dilemma is the class of conservative politicians in countries
such as Germany and Italy in which embryo research is strictly prohibited by law, but
in which hESC research is nevertheless permitt ed or even encouraged, although only
with raw material imported from countries with more permissive laws. For these politicians,
complicity is, and should be, the stumbling block lying in the way of pragmatic
compromise”.
The potential for using PSCs (e.g. hESCs and induced pluripotent stem cells (iPSCs) for
specifi c diseases and conditions is reviewed in the next 16 chapters. Heart disease is
the biggest killer in the Western world (USA - 26% of all deaths), and in the next section,
‘Tissue-specifi c Regeneration of the Heart’, Hao et al. examine the possibility of
one day engineering autologous replacement cardiomyocytes via iPSCs, and review
the small molecules that to date direct diff erentiation of stem cells down the cardiomyogenic
pathway. Sasaki and Okano next discuss the advantages and disadvantages
of diff erent methods for the generation of cardiomyocytes from ESCs – namely the
hanging drop method, suspension culture method and the cell-patt erning method.
X Preface
They end their chapter with a discussion of the cell sheet method for transplantation of
cardiomyocytes into the patient.
Age-related neurodegeneration leading to stroke, dementia and Parkinson’s disease accounts
for around 10% of deaths in the United States. In the Section on ‘Tissue-specifi c
Regeneration of the Brain and Sensory Organs’, Kujoth and Başkaya report on the use
of ESC-derived multipotent mesenchymal stromal cells (MSC; also known as mesenchymal
stem cells) for the treatment of stroke from studies of focal ischemia in the rat.
These authors discuss the mechanisms by which MSC may be trophic, as well as the
potential pitfalls - allograft rejection and limited long-term cell survival/replacement –
in the hostile post-ischemic environment, issues that likely pertain to all replacement
therapies in all tissues of the aging individual. In the next chapters, Fan et al. discuss
the use of ESC transplantation for a range of neurological diseases, while Daadi focuses
on the cellular and molecular control of neural stem cell derivation from adult
and pluripotent stem cells and their diff erentiation into dopaminergic lineage for the
treatment of Parkinson’s disease. Hargus and Bernreuther next weigh in on the practical
issues that need to be overcome in order to successfully transplant neurons into
the brain in order to restore function. As the authors elaborate, these factors include
adequate diff erentiation, survival, migration, and integration of transplanted cells, as
well as the prevention of teratomas. The authors next provide examples, including
from their own research, of how cell adhesion molecules (L1) and extracellular matrix
molecules (tenascin-R) can be applied (transfected) to successfully modify ESCs for cell
therapy approaches in animal models of neurological diseases. These molecules provide
important support to cells, participate in the control of cell development (neurite
outgrowth, synapse formation, and cell migration), and mediate cell survival both in
vitro and in vivo. Neuron support and surveillance also is provided by microglia, and
in the next chapter, Roy et al. describe protocols for the diff erentiation of human and
mouse PSCs into microglia, and how such cells can be expanded in number for drug
screening and cell therapies (combating cancer, neurodegeneration and repair of brain
lesions).
Familial and age-related sensory loss (that aff ect nearly all with age) is the focus of the
next 3 chapters. Sensorineural hearing loss is a major public health problem caused
by the loss or damage of sensory hair cells in the organ of Corti and degeneration of
spiral ganglion neurons (SGNs) or neurons in the auditory brainstem. Hashino and
Fritsch and Nishimura report on the present status of development of stem cell-based
therapies aimed at inner ear regeneration. Importantly, Hashino and Fritsch describe
the how T cell leukemia 3 (Tlx3) confers ESCs diff erentiation into neurons with a glutamatergic
neurotransmitt er phenotype, which is accompanied by establishment of
proper synaptic assembly and axon outgrowth. The authors also highlight the fact that
Tlx3-expressing ESCs can migrate towards degenerating SGNs in the inner ear of host
animals and how such cells may be used to replace damaged SGNs, which cause irreversible
hearing loss in humans. Both authors discuss technical issues related to surgical
approaches for safe and effi cient transplantation of stem cells in the human cochlea,
as well as, non-invasive monitoring of stem cell engraft ment in the cochlea. The potential
use of PSCs for the treatment of retinal diseases such as diabetic retinopathy and
age-related macular degeneration are examined in the next chapter by Messina, who
examines current protocols (and hurdles) for the diff erentiation of mouse iPSCs into
photoreceptors and the functional integration of the transplanted cells. As suggested
Preface XI
by the author, taking cues from the hormonal signals regulating retinal development
and diff erentiation will help improve in vitro protocols for photoreceptor production.
The tissues with the most immediate potential for replacement are various hematopoietic
cell types. In the next 3 chapters of the section ‘Tissue-specifi c Regeneration of Hematopoietic
Systems’, Feng et al., Yokoyama et al. and Nakamura report on the various
methods that have been used to generate almost all types of blood cells from hESCs
(and mESCs), including functionally mature erythrocytes and neutrophils, platelets,
megakaryocytes, eosinophils, monocytes, dendritic cells (DCs), nature killer (NK) cells,
mast cells, and B-, T-lineage lymphoid cells. As the authors note, these advances will
surely translate into clinical applications in the short-term in the fi eld of transfusion
therapies (erythrocytes and platelets) and immune therapies (NK cells and DCs).
Regenerative medicine approaches are described for the liver, thymus and gonads in
the next three chapters of the section ‘Tissue-specifi c Regeneration of Other Tissues’.
Imamura describes research on the generation of hepatocytes for liver disease; the culturing
of embryoid bodies in collagen scaff olds for 24 days with exogenous growth
factors and hormones associated with liver development results in cord-like structures
containing immature hepatocytes. They demonstrate that these cells within collagen
scaff olds form hepatic lobule-like aggregates in the livers of partially hepatectomized
mice. Similarly, Lai demonstrate that murine ESCs can be selectively induced to diff erentiate
into thymic epithelial progenitors with specifi c growth factors. Age-dependent
thymic involution, various genetic and infectious diseases, and protracted T cell defi -
ciencies following chemotherapy or radiotherapy and preparative regimens for foreign
tissue or organ transplants, are some of the conditions that would benefi t from this
research. Kerkis et al. next describe the remarkable progress that has been made in the
diff erentiation of mouse ESCs in the derivation of germ cells and male and female gametes,
although further research is required to generate functional gametes that could be
used for reproductive applications.
In the fi nal section of this book, ‘Side Eff ects of Pluripotent Stem Cell Therapies’, 2
groups review the potential of pluripotency for normal tissue diff erentiation, but the
consequences of the loss of ESCs genome stability. Li and Tanaka review how intrinsic
and extrinsic factors can promote the uncontrolled diff erentiation of PSCs leading
to tumorigenesis, while Moriguchi et al. fi nd that induction of p21 is necessary to
avoid malignant transformations of human iPSCs. Silván et al. examine the similarities
between cancer stem cells and ESCs and the potential for the Yamanaka genes to
induce cancer stem cells. Finally, Rebuzzini et al. point out that propagation of ESCs
for extended culture periods leads to recurrent abnormalities in hESCs and random
karyotypic changes in all chromosomes, changes that must be continually monitored.
These authors describe chromosome abnormalities in ESC lines of human, primates
and rodents and the possible causes of karyotype variation during culture.
It is hoped that the research and reviews described here will help to update the ESC research
community on recent advances in the generation of tissue specifi c cell types for
regenerative applications. The next few decades will see the realization of the potential
for hESCs to treat certain diseases and conditions. However, it is clear from these
reviews that much progress is required in the areas of PSC diff erentiation into tissuespecifi
c cells, transplantation and immune rejection, and genomic stability before PSCs
XII Preface
can fulfi ll their promise. While certain small FDA clinical trials are currently underway,
if the history of another ‘Cinderella’ technology, gene therapy, is any marker of
the progression of PSC research for clinical treatment, it will likely be decades before
PSCs are routinely used to treat age-related diseases, with perhaps the exception of
hematopoietic applications.
References
Thomson JA, Itskovitz-Eldor J, Shapiro SS, Waknitz MA, Swiergiel JJ, Marshall VS &
Jones JM 1998 Embryonic stem cell lines derived from human blastocysts. Science 282
1145-1147.
Craig S. Atwood
Geriatric Research, Education and Clinical Center,
Veterans Administration Hospital,
Department of Medicine,
University of Wisconsin,
Madison, WI 53705, USA