Increasing evidence suggests that liver stem cells have the capacity to differentiate into parenchymal hepatocytes or into bile ductular cells. These stem cells may be activated to proliferate after severe liver injury or exposure to hepatocarcinogens. Stem cell replacement strategies are being investigated as an alternative approach to liver repair and regeneration. Additionally, stem cell transplantation has been shown to significantly improve liver function and increase survival in experimentally-induced liver-injury models in animals. In Liver Stem Cells: Methods and Protocols, expert researchers focus on several hepatic progenitor cells, hepatic differentiation form stem cells, bile ductal cell formation from stem cells, liver stem cells and hepatocarcinogenesi, and application of liver stem cells for cell therapy. These topics shed light on stem cell technology which may lead to the development of effective clinical modalities for human liver diseases. Written in the highly successful Methods in Molecular Biology (TM) series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and key tips on troubleshooting and avoiding known pitfalls. Thorough and intuitive, Liver Stem Cells: Methods and Protocols seeks to aid scientists in the further study of preclinical and clinical investigations that explore the therapeutic potential of stem cells in repair of liver injuries.

Recent stem cell research has revealed that miRNA and RNAi-mediated gene regulation is one of the vital determinates controlling the state of cell differentiation, with the small RNAs serving as key elements involved in regulatory network control of pluripotent cell fate determination. In RNAi and microRNA-Mediated Gene Regulation in Stem Cells: Methods, Protocols, and Applications, expert authors from laboratories across the globe contribute an accessible compendium of up-to-date, proven methods focused on the study of the titular topic. Divided into three sections, the book first gives a brief introduction to RNAi and miRNAs in stem cells, with a focus on the current status of research and future perspectives, then it continues with detailed methods and protocols for RNAi screening, transfection, and the knockdown of specific genes and pathways in several animal species, including humans and mice, concluding with a section on recently developed methods for identification of miRNAs, including a general protocol for preparation and analysis of miRNA libraries for deep sequencing, knock down of a specific gene using miRNA-based shRNA, and miRNA expression analysis using qRT-PCR. Written in the highly successful Methods in Molecular Biology (TM) series format, chapters contain introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and notes highlighting tips on troubleshooting and avoiding known pitfalls. Authoritative and cutting-edge, RNAi and microRNA-Mediated Gene Regulation in Stem Cells: Methods, Protocols, and Applications serves as a valuable resource for scientists and aspiring graduate students interested in the intersection of RNAi, miRNA, and stem cell molecular biology and the exciting areas of medicine, including regenerative medicine, aging, cancer, and neurological disorders, that can be advanced through this expanding area of research.

Stem cell mobilization represents a transient increase in the levels of circulating stem and progenitor cells. In Stem Cell Mobilization: Methods in Protocols, expert researchers in the field detail cell mobilization methodology and recent developments in the field for basic and biomedical research community. Specifically clinical hematopoietic progenitor cell mobilization protocols, frontiers in mobilization and analysis of non-hematopoietic progenitors, mesenchymal progenitor cells, monocyte-derived fibroblast progenitors, and very small embryonic like cells. Written in the highly successful Methods in Molecular Biology (TM) series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and key tips on troubleshooting and avoiding known pitfalls. Authoritative and practical, Stem Cell Mobilization: Methods and Protocols seeks to aid scientists in the further study of this important stem cell research.

In 2010 the global area of transgenic crops reached 148 million hectares, an 87-fold increase since 1996, making it the most rapidly adopted technology in the history of modern agriculture. In Transgenic Plants: Methods and Protocols, Second Edition expert researchers in the field provide key techniques to investigate production and analysis of transgenic plants. Focusing on selection and detection methods, transformation technology, gene targeting, silencing and directed mutation, metabolic engineering and pharming, the book encompasses protocols relating to major crops and model plants being used for genomic analysis. Written in the highly successful Methods in Molecular Biology (TM) series format, the chapters include the kind of detailed description and implementation advice that is crucial for getting optimal results in the laboratory. Thorough and intuitive, Transgenic Plants: Methods and Protocols, Second Edition aids scientists in the continuous improvements being made for the production and analysis of transgenic plants.

The migration of stem cells has been found to be critical during early development for the organization of the embryonic body as well as during adult life with tissue homeostasis and regeneration of organ function. Due to the low frequency of these cells in vivo, problems in identifying and prospectively purifying tissue specific stem cells near homogeneity, and, most importantly, a lack of adequate technologies and protocols to study stem cell migration in vivo, this vital research has been quite difficult until recently. In Stem Cell Migration: Methods and Protocols, experts in the field compile and highlight the standard and novel techniques that allow the studying of the migration of stem cells in one succinct manual. Including protocols on germ, neuronal, and hematopoietic stem cells, during development and adulthood with a clear emphasis on in vivo technologies, the volume also extends its coverage to in vitro approaches toward several developmentally-conserved signaling pathways. Written in the highly successful Methods in Molecular Biology (TM) 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. Practical and convenient, Stem Cell Migration: Methods and Protocols provides key, state-of the art information on experimental techniques for studying stem cell migration both at a cellular and molecular level in development, regeneration, and disease.

Over time, it has become clear that changes in stem cells do occur during aging, not only in their number but also in their relationship to their microenvironment and their functionality as reflected in changes to their metabolome. Stem Cells and Aging: Methods and Protocols brings together chapters from expert contributors with protocols critical for exploring the biology of stem cell aging, all of which is key for understanding these age-related stem cell changes at a basic biology level and at the level of their impacts for regenerative medicine. Written in the highly successful Methods in Molecular Biology (TM) series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible protocols, and tips on troubleshooting and avoiding known pitfalls. Concise and easy to use, Stem Cells and Aging: Methods and Protocols serves as an ideal reference to guide investigators toward further valuable answers to the problems of our aging population.

Stem Cell Transcriptional Networks: Methods and Protocols collects techniques used to increase our understanding of the underlying transcriptional programs of stem cells that promote self-renewal and differentiation. The volume opens with a section on next-generation sequencing library preparation and data analysis. Continuing with a collection of protocols on visual analysis and interpretation of large-scale interaction networks, this detailed compilation features transcriptional networks in embryonic and adult stem cells, embryo culture and derivation of stem cells, as well as transcriptional programs that promote self-renewal, reprogramming, and transdifferentiation. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials, step-by-step, readily reproducible protocols and tips on troubleshooting and avoiding known pitfalls. Authoritative and practical, Stem Cell Transcriptional Networks: Methods and Protocols aims to provide a key resource for biologists seeking to interrogate these vital networks.

In Epiblast Stem Cells: Methods and Protocols, expect researchers in the field provide a detailed collection of techniques and protocols useful to the study of the biology of the pluripotent epiblast. These include methods and techniques used to study epiblast development in different amniotes. This collection brings together contributions from the fields of embryology, stem cell biology and developmental biology together, providing a single volume with detailed procedures for the isolation and culture of epiblasts at different stages of development, and techniques for the study of differentiation into specific lineages. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, a complete list of the necessary materials and reagents, detailed laboratory protocols, and extensive notes providing suggestions on troubleshooting and how to overcome common difficulties. Comprehensive and cutting-edge, Epiblast Stem Cells: Methods and Protocols serves as a resource to individuals interested in studying the biology of pluripotent cells.

Mapping of animal genomes has generated huge databases and several new concepts and strategies, which are useful to elucidate origin, evolution and phylogeny. Genetic and physical maps of genomes further provide precise details on chromosomal location, function, expression and regulation of academically and economically important genes. The series Genome Mapping and Genomics in Animals provides comprehensive and up-to-date reviews on genomic research on a large variety of selected animal systems, contributed by leading scientists from around the world. Laboratory animals are those species that by accident of evolution, domestication and selective breeding are amenable to maintenance and study in a laboratory environment. Many of these species are studied as 'models' for the biology and pathology of humans. Laboratory animals included in this volume are sea-urchin, nematode worm, fruit fly, sea squirts, puffer fishes, medaka fish, African clawed frog, mouse and rat.

Mesenchymal Stromal Cell (MSC) biology has been studied for more than 4 decades and the cells have been investigated for potential clinical applications for more than 15 years. Progress has become exponential over the past decade due mainly to the broad therapeutic potential of these cells. However, MSC studies have also been subject to controversy and increasing scrutiny as new mechanisms of action are reported and ever-expanding therapeutic applications pursued. In this book, leading authorities from all over the world, who are actively involved in this field, provide state-of-the-art knowledge of the basic biology, translational requirements and latest clinical experience with MSCs. This cutting edge book is the ideal resource for scientists and clinicians interested in pursuing an important and rapidly developing field of research that will eventually help patients and address urgent unmet medical needs. Features include: Coverage of the biology of MSCs, latest understanding of mechanisms of action, and role in tissue homeostasis and regeneration Identifying the potential of MSCs in proceeding from bench to bedside from regulatory , GMP production, ethical and safety aspects Critical analysis of clinical studies and the potential of MSCs to treat a wide variety of human diseases and tissues.

Stem cells are found in almost all organisms from the early stages of development to the end of life. There are several types of stem cells and all of them may prove useful for medical research; however, each of the different types has both promise and limitations. Somatic Stem Cells: Methods and Protocols presents selected genetic, molecular, and cellular techniques used in somatic stem cell research and its clinical application. Chapters focus on the isolation, characterization, purity, plasticity, and clinical uses of somatic stem cells from a variety of human and animal tissues. Written in the highly successful Methods in Molecular Biology (TM) series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and key tips on troubleshooting and avoiding known pitfalls. Through and intuitive, Somatic Stem Cells: Methods and Protocols seeks to provides scientists with the fundamental techniques of stem cell research and its potential application in regenerative medicine.

Before the therapeutic potential of cell replacement therapy or the development of therapeutic drugs for stimulating the body's own regenerative ability to repair cells damaged by disease and injury can be fully realized, control of stem cell fate, immuno-rejection, and limited cell sources must be overcome. In Cellular Programming and Reprogramming: Methods and Protocols, expert researchers cover the most recent technologies and their related mechanisms involved in the programming and reprogramming of cell fate. Written in the highly successful Methods in Molecular Biology (TM) series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, laboratory protocols, and notes to highlight tips on troubleshooting and avoiding known pitfalls. Essential and cutting-edge, Cellular Programming and Reprogramming: Methods and Protocols promises to aid scientists attempting to advance stem cell biology in order to better treat devastating human diseases, including cardiovascular disease, neurodegenerative disease, musculoskeletal disease, diabetes, and cancer.

Stem Cell therapy for lysosomal diseases (LSDs) is developing rapidly. This volume discusses the history, current practice and future perspectives of stem cells in inborn errors of metabolism (IEM) and provides an international perspective on progress, limitations, and future directions (e.g. gene therapy, iPS, ES) in the field. Beginning with an overview of these diseases, the book covers the breadth of this topic from treatment options, bone marrow transplantation, and alternative treatment options, through long-term outcomes and future perspectives.

Progenitor cells have become important in regenerative medicine therapies, due to their potential to differentiate into many cell types. This capability, and understanding how to regulate the cells, will provide the basis for future cell therapies aimed at correcting tissue and organ dysfunction as a result of disease or injury. In, Progenitor Cells: Methods and Protocols, expert researchers in the field detail many of the methods which are now commonly used to investigate progenitor cells. These include methods and techniques of the manipulation of physical forces that shape progenitor cell behavior, studying progenitor cells in vivo, using non-mammalian and mammalian model systems, and investigating human progenitor cells, including their isolation, characterization and application in cell-based therapies. Written in the highly successful Methods in Molecular Biology (TM) series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and key tips on troubleshooting and avoiding known pitfalls. Authoritative and practical, Progenitor Cells : Methods and Protocols seeks to aid scientists in the further study progenitor cells and how they are studied across multiple systems.

This text highlights the endogenous regenerative potential of the central nervous system in neonates and juveniles and discusses possible ways it might be manipulated for medical purposes. The first section provides a descriptive summary of the salient steps of human brain development with a discussion of comparisons with other mammalian brains. It also provides a historical perspective on our understanding of ongoing brain development throughout the lifespan and serve to introduce the concept of brain plasticity following injury. The second part is devoted to the endogenous reparative potential of the brain, including its limitations, and articles focusing on defined pathologies (e.g. anoxia/hypoxia, epilepsy, traumatic brain injury and stress) in animal models and in humans pinpoint eventual ways these pathologies might be manipulated. The third and final focuses on the "dark side" of stem cells for brain repair or of the manipulation of spontaneous adaptive events after injury (e.g. genomic instability, sensitization to cancerous transformation and defective neural networks).

Almost daily, new technologies are being presented that move the field of human pluripotent stem cell research towards a future that may yield highly-effective, personalized medical treatments. Three enabling technologies at hand for human PSCs are 1) directed reprogramming of somatic cells, which eliminate many of the ethical issues associated with the derivation and use of human PSCs, increase genetic diversity of the available human PSC lines, and give rise to better in vitro human disease models; 2) the discovery that a Rho-associated protein Kinase (ROCK) inhibitor allows for efficient single cell passaging and cryopreservation, increasing the efficiency and reliability of hPSC culture; and 3) defined, animal-component-free media, which lay the groundwork for simplified scale-up for therapeutic applications, differentiation protocols, and toxicology screens. The aforementioned technologies can be found in Human Pluripotent Stem Cells: Methods and Protocols, a compilation of 33 detailed protocols in six categories of PSC research that cover laboratory essentials and the derivation of new PSC lines, including induced PSC lines, as well as their growth, maintenance, characterization, genetic manipulation, and differentiation. Written in the successful Methods in Molecular Biology (TM) series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible protocols, and notes on troubleshooting and avoiding known pitfalls. Authoritative and accessible, Human Pluripotent Stem Cells: Methods and Protocols serves as an ideal guide to scientists conducting their own pluripotent cell research programs and makes great strides towards furthering human knowledge and, ultimately, improving the human condition.

Considerable advances have taken place since the initial isolation and characterization of human embryonic stem (HES) cells; however, significant challenges remain before their potential for restoration and regeneration processes in patients can be realized. Understanding the diversity amongst HES cell lines and realizing the ability to isolate lines with robust differentiation potential remain difficult. In the Human Embryonic Stem Cells Handbook, experts in the field provide an assortment of protocols that have been used by various laboratories around the world so as to allow both novices and experienced investigators to compare and contrast different approaches to HES cell isolation and characterization with the hope that, from these protocols, researchers might standardize approaches for HES cell biology. Written in the Methods in Molecular Biology (TM) 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 for troubleshooting and avoiding known pitfalls. Authoritative and accessible, Human Embryonic Stem Cells Handbook serves as a valuable reference for scientists pursuing this vital field and its enormous potential.

This timely volume explores tissue-derived stem cells, summarizing and evaluating the current body of knowledge. Sixteen chapters cover hematopoietic, osteoblast, skeletal, cardiac, pancreatic, epithelial, corneal, hepatic, renal, testicular, ovarian, fingernail, hair follicle and other stem cells- discussing their individual development, functions and pathologies, as well as the commonalities and interdependence between niches. This installment of the popular Stem Cell Biology and Regenerative Medicine series delivers authoritative, international perspectives on this rapidly growing field. Tissue-Specific Stem Cell Niche is an ideal complementary volume to Biology of Stem Cell Niche and Adult Stem Cells, Second Edition and will be invaluable to clinicians and researchers working with stem cells as well as to postgraduate trainees who are studying them.

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.

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.

With the discovery of stem cells capable of multiplying indefinitely in culture and differentiating into many other cell types in appropriate conditions, new hopes were born in repair and replacement of damaged cells and tissues. The features of stem cells may provide treatment for some incurable diseases with some therapies are already in clinics, particularly those from adult stem cells. Some treatments will require large number of cells and may also require multiple doses, generating a growing demand for generating and processing large numbers of cells to meet the need of clinical applications. With this in mind, our aim is to provide a book on the subject of stem cells and cell therapy for researchers and students of cell biotechnology, bioengineering and bioproduction. This book is exceptional as it teaches researchers stem cells and cell therapy in that it covers the concepts and backgrounds necessary so that readers get a good understanding of the production of stem cells. The book covers three topics: The basics of stem cells and cell therapy, the use of stem cells for the treatment of human diseases, and stem cell processing. It includes chapters on neural and vascular stem vascular stem cell therapy, expansion engineering of embryonic stem cells, stem cell based production of blood cells and separation technologies for stem cells and cell therapy products. It is an informed and informative presentation of what modern research, science and engineering have learned about stem cells and their production and therapies. Addressing both the medical and production issues, this book is an invaluable contribution to having an academic and industrial understanding with respect to R&D and manufacturing of clinical grade stem cells.

This comprehensive volume explores functions, pathologies, and applications of stem cells in relation to the niches in which they develop. Ten chapters cover the subject in depth, from a historical perspective through signaling, hormonal control, quiescence, biomimetics, epigenetics, engineering strategies for emulating, tumorigenesis and more. The chapter authors represent a broad range of international expertise and perspectives. This installment of the popular Stem Cell Biology and Regenerative Medicine series delivers authoritative, international perspectives on this rapidly growing field. Biology of Stem Cell Niche is an ideal complementary volume to Tissue-Specific Stem Cell Niche and Adult Stem Cells, Second Edition and will be invaluable to clinicians and researchers working with stem cells as well as to postgraduate trainees who are studying them.

“Perspectives in Inflammation Biology” outlines detailed studies using preclinical murine models in Inflammation. The book is meant for academicians, industry persons, research scholars and students alike. The detailed perspective for a beginner and the exhaustive methodologies and analyses outlined, for the veteran researcher, makes this book a unique link between someone who is thinking of embarking on a study of inflammation and one who is delving deep into this area of specialization. The book deals with asthma and allergy as specific disease areas of inflammation of the lung, aseptic peritonitis as a disease of systemic inflammation and details how each role player in its pathophysiology has a unique niche of activity. Data acquisition, sequentiality and analyses in context demonstrate how each role player is validated systematically to become a target for drug discovery. Methods and models used in the course of my work and their relevance will demonstrate to the researcher how a study can be developed from an idea. Further into a researcher’s ongoing work, this book is meant to stimulate new questions and pave ways for better dissection of a phenomenon. The highlights of this book are the detailed tables tabulating sub species of immune cells, their inflammatory recruitment indices, their translation into tissue-to-tissue traffic of the inflammatory stimulus and the delicate interplay of resident structural cells, cells recruited from circulation, their feedback poiesis in bone marrow, their instruction in the lymphoid organs and tissues as well as the non-cellular mediators synthesized from corresponding genetic instruction. The book shall definitely help students and researchers how a disease can be simplified from its complex ramifications and network of implications and put back into perspective and the whole thing falls into place without an intimate understanding of the mechanism and the compelling circumstances that causes a disease, a drug hunter cannot hope to begin her quest. To find the “Achilles’ heel” and effectively neutralize the enemy!

This is the thirteenth volume in a series which presents current understanding of stem cells and cancer stem cells. This book presents many relevant topics including the different sources of cancer stem cells: the features, roles and clinical potential of stem cells emerge from engaging with this comprehensive work. Chapters explore molecular mechanisms underlying human somatic cell reprogramming to generate induced pluripotent stem cells and provide expert discussion of the advantages and limitations of applications of some of the stem cell types (pluripotent stem cells, neural stem cells). Many advances are explored, including the importance of stem cell markers in diagnosis, the role of stem cells in angiogenesis and a method for isolating multi potent endothelial-like cells from human adipose tissue, to name a few. Readers will also find a consideration of mathematical models and other quantitative tools which could facilitate research and discovery in the field. The editor's preface to the book and the series is particularly helpful in introducing the work presented in this volume. The detailed overviews, practical experience and insights of international expert authors presented in this handbook will be of value to both professional practitioners and scholars in basic research.

Bone marrow stem cells are the most transplanted cells worldwide. These cells are used as a replacement therapy for patients suffering from a diverse number of hematopoietic diseases and immunodeficiencies. However, the use of bone marrow cells in regenerative medicine has so far remained without much success. In the new era of pluripotent stem cells, great opportunities for establishing new therapies have opened up. The discovery of human embryonic stem cells and that of induced pluripotent (iPS) stem cells has made it possible to derive any desired tissues for regenerative medicine as iPS cell derived cells are only limited by the lack of established protocols that can be applied in humans. There is no doubt that stem cells present a new and innovative platform for establishing novel cell based therapies. The challenge is to establish new protocols that allow the successful differentiation of these cells into lineage committed cells. Embryonic Stem Cell Immunobiology: Methods and Protocols covers a variety of relevant topics, such as hematopoietic stem cells derived from ES cells, the interaction of these cells with natural killer cells or with cytotoxic T cells, and specific protocols for the derivation of hematopoietic cells and neuronal cells, to name a few. Written in the highly successful Methods in Molecular Biology series format, chapters contain introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and notes on troubleshooting and avoiding known pitfalls. Authoritative and accessible, Embryonic Stem Cell Immunobiology: Methods and Protocols serves as an ideal guide to experts and non-experts interested in different aspects of stem cells.