This volume presents up-to-date methods that allow primary stem cells from a variety of sources to be isolated, cultured in vitro, detected and measured for specific applications. These applications range from those in basic, stem cell and veterinary research to toxicology, cellular therapy and regenerative medicine. There is a slight bias towards the blood-forming system as more is known about the blood-forming or hematopoietic system than any other primary stem cell system. These unique properties and characteristics are discussed and examined, mostly at the cellular level and in detail in this book. Written in the 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 protocols, and notes on troubleshooting and avoiding known pitfalls. Authoritative and accessible, Stem Cell Protocols provides novices with the fundamentals necessary to develop new technologies necessary for basic and clinical research in the future, and will aid professionals in finding new methodologies to provide a wider viewpoint and an even greater scope for their own research.

This book presents a comprehensive overview of the current understanding of the organization of endogenous lung stem and progenitor cell compartments during fetal lung development, postnatal lung growth and in adulthood. Progressing stage by stage, the chapters on fetal lung development emphasize the integrated role of epithelial, stromal, vascular and neural cell elements in building a functional lung, while the subsequent chapters on adult lung regeneration describe the nature and properties of adult lung stem/progenitor cells distributed along the proximal-distal axis of the airway tree. The chapters on regulation of lung regeneration and repair discuss how regenerative cells interact with their niche microenvironment and how regulation of lung regeneration and repair in the steady state and following injury recapitulates ontogeny. And, lastly, the chapters on cellular therapies for lung disease and bioengineering the lung focus on promising emerging therapies and approaches in lung regenerative medicine. The scope of this volume of the Stem Cell Biology and Regenerative Medicine series focuses on exploring the topic of building and rebuilding the lung from a cellular rather than a molecular perspective. Thus, the section on cellular therapies does not include extensive coverage of each of the numerous lung diseases, including cancer, which could be amenable to stem cell-based therapies, although the final chapter does include some discussion on the future prospects and challenges. All of the contributors are working on the cutting edge of the lung stem cell field, making this book essential reading for those with an interest in the field of lung stem cell biology and the potential role of cellular therapies and tissue bioengineering approaches in lung regenerative medicine, including biomedical scientists, graduate students, post-graduate researchers and respiratory clinicians.

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.

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.

Biomimetics and Stem Cells: Methods and Protocols collects a series of approaches to demonstrate the role and value of biomimetics for the better understanding of stem cell behavior and the acceleration of their application in regenerative medicine. Recent advances in tissue engineering are enabling scientists to ''instruct'' stem cells toward differentiating into the right phenotypes, in the right place and at the right time. Given these advances, biomimetic environments are being designed to recapitulate, in vitro, the combinations of factors known to guide tissue development and regeneration in vivo and thereby help unlock the full potential of the stem cells. Written in 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. Practical and essential, Biomimetics and Stem Cells: Methods and Protocols focuses on the use of biomimetic systems for stem cells in order to aid in moving this vital field of study forward.

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.

This book represents a major contribution to the emerging science of regenerative medicine using non-fetal sources of stem cells. The Editors, Dr Niranjan Bhattacharya and Professor Phillip Stubblefield, have brought together some of the most pre-eminent scientists working on regenerative medicine to share information on currently ongoing work in this area alongside unpublished observations that will help to shape the contours of future therapies. Regenerative Medicine: Using Non-Fetal Sources of Stem Cells discusses the potential clinical and therapeutic applications using non-fetal stem cells as well as providing instruction on the collection, isolation and characterization of stem cells from various non-fetal sources, such as menstrual blood, adipose tissue, breast milk and uprooted decidual teeth. This book will be an invaluable resource for both active researches and those entering the field. The Editors truly hope that the text will act as a stimulant to professionals and clinical scientists, who may be inspired to further the work of the pioneering scientists who have contributed to this volume.

The ability to produce vast amounts of DNA sequence data has enabled the discovery of molecular markers in model organisms, crops, as well as orphan species making genotyping the rate limiting factor, and this volume focuses on the different markers available and the low to high throughput genotyping of these markers. Given the diverse nature of some of these systems, an overview is provided on the identification of markers from sequence data, as well as data analysis with example applications once the genotyping data has been generated. Written in the 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 protocols, and notes on troubleshooting and avoiding known pitfalls. Authoritative and easily accessible, Plant Genotyping: Methods and Protocols is aimed at plant molecular biologists, geneticists, plant breeders and ecologists who have a target question and need to know the most suitable markers and genotyping system to use.

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.

The discovery of stem and progenitor cells in the adult mammalian CNS challenged the long standing "no new neuron" doctrine and opened the door to the potential for cell replacement therapy. The process from discovery to clinical applications can be long and tortuous, requiring rigorous steps involving standardized and precise protocols. Neural Progenitor Cells: Methods and Protocols is a collection of practical articles describing techniques used to study neural stem and progenitor cells. The volume also highlights the promise of stem cell-based therapeutic applications for CNS disorders. Written in the 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 protocols, and notes on troubleshooting and avoiding known pitfalls. Authoritative and easily accessible, Neural Progenitor Cells: Methods and Protocols serves both professionals and novices with its well-honed methodologies.

Rhim and Kremer's state-of-the-art volume on Human Cell Transformation: Role of Stem Cells and the Microenvironment highlights the latest findings on the current state of human cell transformation model systems and provides the insight into the molecular and cellular changes involved in the conversion of normal cells to neoplastic cells. Chapters cover all recently developed novel human cell models. In addition, the rapidly growing fields of knowledge regarding not only stem cells in cancer progression, but also the role of the microenvironments in human carcinogenesis are discussed. A wealth of topics is presented including: * Derivation of epithelial, fibroblastic, and hematopoietic in vitro model systems * Oncogenes * Tumor suppressor genes * Viral transformation * In vitro model systems for viral, chemical and radiation carcinogenesis * Cell aging * The multistep nature of human carcinogenesis * The role of stem cells and the microenvironment in tumorigenesis * The genes involved in multistep carcinogenesis Unique in both scope and focus - devoted solely to human cell transformation systems - Human Cell Transformation: Role of Stem Cells and the Microenvironment provides unparalleled, in-depth coverage for cancer researchers, cell and molecular biologists, hematologists, virologists, and workers in related fields. Essential reading for everyone who needs to be kept up-to-date in this fast-paced area! Features O Multistep models O Breast cancer/Stem cells O Prostate cancer/Stem cells O Multistep / Genes

Interest in a specialized microenvironment or "niche" regulating hemopoietic stem cell function has been steadily growing since the idea was first proposed by Ray Schofield over three decades ago. This growing interest, as well as more recently the interest in cellular-molecular-biochemical characterization of not only the hemopoietic stem cell niche but the niches for other stem cells, incited the compilation of Stem Cell Niche: Methods and Protocols. In this volume, scientists have provided protocols that will provide both a flavor of the field and hopefully stimulate new approaches and methodologies by those interested in the stem cell niche. Written in the 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 protocols, and notes on troubleshooting and avoiding known pitfalls. Authoritative and easily accessible, Stem Cell Niche: Methods and Protocols seeks to serve both experts and novices in the stem cell field with well-established protocols on this exciting subject.

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.

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.

“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 book offers readers cutting-edge research at the interface of polymer science and engineering, biomedical engineering, materials science, and biology. State-of-the-art developments in microscale technologies for cell engineering applications are covered, including technologies relevant to both pluripotent and adult stem cells, the immune system, and somatic cells of the animal and human origin. This book bridges the gap in the understanding of engineering biology at multiple length scale, including microenvironmental control, bioprocessing, and tissue engineering in the areas of cardiac, cartilage, skeletal, and vascular tissues, among others. This book also discusses unique, emerging areas of micropatterning and three-dimensional printing models of cellular engineering, and contributes to the better understanding of the role of biophysical factors in determining the cell fate. Microscale Technologies for Cell Engineering is valuable for bioengineers, biomaterial scientists, tissue engineers, clinicians, immunoengineers, immunologists and stem cell biologists, as it offers a review of the current cutting-edge cell engineering research at multiple length scale and will be valuable in developing new strategies for efficient scale-up and clinical translation.

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.

Human pluripotent stem cells such as human embryonic stem cells (hESC) and induced pluripotent stem cells (iPSC) with their unique developmental plasticity hold immense potential as cellular models for drug discovery and in regenerative medicine as a source for cell replacement. While hESC are derived from a developing embryo, iPSC are generated with forced expression of key transcription factors to convert adult somatic cells to ESC-like cells, a process termed reprogramming. Using iPSC overcomes ethical issues concerning the use of developing embryos and it can be generated from patient-specific or disease-specific cells for downstream applications. Pluripotent Stem Cells: Methods and Protocols highlights the best methods and systems for the entire work flow. Divided into four convenient sections, topics include a focus on producing iPSC from diverse somatic sources, media systems for expanding ESC and iPSC with detailed protocols for directed differentiation into specific lineages, commonly used cellular and molecular characterization methods , and the potential application of labeled stem cells with specific methods for cloning, gene delivery and cell engineering. 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 easily accessible, Pluripotent Stem Cells: Methods and Protocols seeks to serve both professionals and novices with its well-honed methodologies in an effort to further our knowledge of this essential cellular feature.

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.

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.

Embryonic stem cells (ESCs) offer an unlimited self-renewing capacity, as opposed to the limits of adult stem cells; therefore, ESCs represent an almost bottomless resource for regenerative medicine and tissue engineering approaches. In Embryonic Stem Cell Therapy for Osteo-Degenerative Diseases: Methods and Protocols, accomplished investigators provide detailed descriptions on how to expand ESCs from the most commonly used species ex vivo, i.e. mouse and human, in static culture as well as in controllable bioreactor processes. The thorough and timely volume summarizes the methods that may be used to differentiate these cells along the desired lineage of choice, be it osteoblasts, osteoclasts, or chondrocytes, and consequentially also offers analysis tools for the characterization of resulting cells and evaluation of differentiation effectiveness. 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 expert tips on troubleshooting and avoiding known pitfalls. Dependable and cutting-edge, Embryonic Stem Cell Therapy for Osteo-Degenerative Diseases: Methods and Protocols supplies the tools necessary to allow researchers to carry out critical research needed in order to bring this burgeoning and vitally important field closer to the clinic and to ensure the widespread application of a successful strategy.

In Cereal Genomics: Methods and Protocols, expert researchers provides modern protocols for the analysis and manipulation of cereal genomes. Techniques for isolation and analysis of DNA and RNA from both the vegetative tissues and from the more challenging seeds of cereals are described. Tools for the isolation, characterization and functional analysis of cereal genes and their transcripts are detailed. Methods for molecular screening of cereals and for their genetic transformation are also covered. Written in 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 practical, Cereal Genomics: Methods and Protocols provides a comprehensive resource for those studying cereal genomes.

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.

Human pluripotent stem cells, including human embryonic stem cells and induced pluripotent stem cells, are a key focus of current biomedical research. The emergence of state of the art culturing techniques is promoting the realization of the full potential of pluripotent stem cells in basic and translational research and in cell-based therapies. This comprehensive and authoritative atlas summarizes more than a decade of experience accumulated by a leading research team in this field. Hands-on step-by-step guidance for the derivation and culturing of human pluripotent stem cells in defined conditions (animal product-free, serum-free, feeder-free) and in non-adhesion suspension culture are provided, as well as methods for examining pluripotency (embryoid body and teratoma formation) and karyotype stability. The Atlas of Human Pluripotent Stem Cells - Derivation and Culturing will serve as a reference and guide to established researchers and those wishing to enter the promising field of pluripotent 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.