Since the first successful isolation and cultivation of human embryonic stem cells at the University of Wisconsin, Madison in 1998, there has been high levels of both interest and controversy in this area of research. This book provides a concise overview of an exciting field, covering the characteristics of both human embryonic stem cells and pluripotent stem cells from other human cell lineages. The following chapters describe state-of-the-art differentiation and characterization of specific ectoderm, mesoderm and endoderm-derived lineages from human embryonic stem cells, emphasizing how these can be used to study human developmental mechanisms. A further chapter discusses genetic manipulation of human ES cells. The concluding section covers therapeutic applications of human ES cells, as well as addressing the ethical and legal issues that this research have raised.

This volume covers all aspects of embryonic stem cell differentiation, including mouse embryonic stem cells, mouse embryonic germ cells, monkey and human embryonic stem cells, and gene discovery.* Early commitment steps and generation of chimeric mice* Differentiation to mesoderm derivatives* Gene discovery by manipulation of mouse embryonic stem cells

This volume will cover a series of reviews on stem cells including adult and embryonic stem cells. Speakers were invited to present these talks during the Stem Cell Symposia in fall of 2010, in Samsun, Turkey. Unique aspect of this volume is that it brings a multidisciplinary aspect of stem cells extracted from a symposium.

This fully updated volume explores improved and extended protocols for embryonic stem cell (ESC) research, provided with the most direct and informative methodologies currently available. The book examines how these models for cell lineage and differentiation studies have continued to mature into a critical research workhorse. Written for the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and up-to-date, Embryonic Stem Cell Protocols, Fourth Edition serves as an ideal resource for researching mining the depths of the ESC field.

Stem cells have the ability to differentiate into all types of cells within the body, thus have great therapeutic potential for regenerative medicine to treat complicated disorders, like Parkinson’s disease and spinal cord injury. There will also be many applications in drug development. However, several roadblocks, such as safety issues and low efficiency of pluripotent stem cell (PSC) line derivation need to be resolved before their clinical application. This thesis focuses on these two areas, so as to find methods to overcome the limitation. It covers deriving embryonic stem cells (ESCs) from several different species and reports an efficient system to generate induced pluripotent stem cells (iPSCs), and the first iPSC mice in the world. The results in this thesis confirm that somatic cells can be fully reprogrammed with the four Yamanaka factors. In addition, we have found that the Dlk1-Dio3 region can be a potential molecular marker to distinguish the fully reprogrammed iPSCs from partially reprogrammed ones. All of these results will help improve the safety of PSCs in the clinical applications and increase the current low induction efficiency of their production.

A much-needed primer on the use of laser flow cytometry for stem cell analysis Laser flow cytometry is a powerful tool for rapid analysis of cells for marker expression, cell cycle position, proliferation, and apoptosis. However, no resources specifically address the use of this methodology for the study of stem cells; this is especially important as stem cell analysis involves specialized methods and staining procedures based on specific characteristics such as marker expression, cell size, drug transport, and efflux of the stem cells. Now, this book reviews these procedures, discusses the science behind them, and provides real-world examples to illustrate the usefulness of the methods. It brings together world-class experts in pathology, biophysics, immunology, and stem cell research, who draw upon their extensive experience with the methods and show examples of good data to help guide researchers in the right direction. Chapter coverage includes: Stem cell analysis and sorting using side population Flow cytometry in the study of proliferation and apoptosis Stem cell biology and application Identification and isolation of very small embryonic-like stem cells from murine and human specimens Hematopoietic stem cells—issues in enumeration Human embryonic stem cells: long-term culture and cardiovascular differentiation Limbal stem cells and corneal regeneration Flow cytometric sorting of spermatogonial stem cells Breast cancer stem cells Stem cell marker expression in cells from body cavity fluids This book is an essential resource for all graduate students, practitioners in developing countries, libraries and book repositories of universities and research institutions, and individual researchers. It is also of interest to laboratories engaged in stem cell research and use of stem cells for tissue regeneration, and to any organization dealing in stem cell and tissue regeneration research.

The emerging field of human embryonic stem cell biomedicine crosses many disciplinary boundaries-cell biology, reproductive biology, embryology, molecular biology, endocrinology, immunology, fetal med

International Workshop organised by the Marcel Mérieux Foundation, 21 to 23 June 2000. The debate over the ethical issues raised by stem cell research concerns essentially the practice of taking cells from human embryos and the consequent destruction of the embryo. This work, going to the heart of the controversy over such manipulations, discusses the ethical question of the legal status of the embryo. At the moment when, in France, the bioethics laws have come up for review, questions regarding the statute of the embryo return in the heart of scientific debates. Breakthroughs in the field of embryonic stem cell biology offer a glimpse of the considerable therapeutic possibilities. Research Institutes and Governments, hailed by these new therapeutic perspectives, are attempting to put in place modes of regulation this research that both respond to citizen's aspirations and conform to ethical norms.

Stem cells appear to be fundamental cellular units associated with the origin of multicellular organisms and have evolved to function in safeguarding the cellular homeostasis in organ t- sues. The characteristics of stem cells that distinguish them from other cells have been the fascinating subjects of stem cell research. The important properties of stem cells, such as ma- tenance of quiescence, self-renewal capacity, and differentiation potential, have propelled this exciting ?eld and presently form a common theme of research in developmental biology and medicine. The derivation of pluripotent embryonic stem cells, the prospective identi?cation of multipotent adult stem cells, and, more recently, the induced pluripotent stem cells (popularly called iPS) are important milestones in the arena of stem cell biology. Complex networks of transcription factors, different signaling molecules, and the interaction of genetic and epi- netic events constantly modulate stem cell behavior to evoke programming and reprogramming processes in normal tissue homeostasis during development. In any given cellular scenario, the regulatory networks can pose considerable complexity and yet exert an orderly control of stem cell differentiation during normal development. An aberration in these ?nely tuned processes during development usually results in a spectrum of diseases such as cancers and neurological disorders. Thisunderscorestheimminentneedforamorecompleteunderstandingofmolecular mechanisms underlying the regulatory circuitries required for stem cell maintenance. Overthepast3–5years,adiversegroupofbenchandphysicianscientistshaveprospectively enhanced our knowledge of stem cell biology. These studies are unveiling many unrecognized or previously unknown fundamentals of developmental biology.

Stem cells, characterized by the ability to both self-renew and to generate diff- entiated functional cell types, have been derived from the embryo and from va- ous sources of the postnatal animals and human. The recent advances in stem cell research have led to a better understanding of self-renewal, maintenance, and diff- entiation of both embryonic and somatic stem cells. This has significantly increased our knowledge of cellular and developmental biology in general and will certainly continue to do so for a long time to come. Moreover, given their role in maintaining and replenishing tissues, stem cells represent a potential means of restoring tissue function and thereby treating the root cause of degenerative disease. Therefore, in parallel, we need to improve our cognizance of the challenges involved in applying stem cells in clinical settings. The current chapters highlight both of these aspects: that of understanding the “actual” and that of developing the “possible. ” In recognition of the growing excitement and potential of stem cells as models for both the advancement of basic science and future clinical applications, I felt it timely to edit this book in which forefront investigators would provide new findings for the use of stem cells to study various lineages and tissue types and some app- cations.