Induced Stem Cells

This extensive new edition presents protocols reflecting the great strides made in the study of induced pluripotent stem (iPS) cells. The collection explores new and improved methods for the generation, expansion, and maintenance of iPS cells from different tissue types, characterization of their differentiation pathways along different lineages, and their potential utility in tissue repair and regeneration. Written for the highly successful Methods in Molecular Biology series, 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. Comprehensive and up-to-date, Induced Pluripotent Stem (iPS) Cells: Methods and Protocols, Second Edition aims to arm stem cell biologists, both novice and expert, with invaluable protocols that are currently being used in various laboratories around the world.

This detailed volume presents a series of protocols that are representative of recent developments and improvements in induced pluripotent stem cells (iPS cells) and corresponding human disease models. Reflecting the latest technology for generating induced pluripotent stem cells (iPS cells) and their initial characterization, the book explores techniques invaluable both for studies of disease-specific cell types and for their potential applications in regenerative medicine. Written for the highly successful Methods in Molecular Biology series, chapters include introduction to their respective topics, lists of the necessary materials and reagents, step-by-step and readily reproducible laboratory protocols, as well as tips on troubleshooting and avoiding known pitfalls. Authoritative and practical, Induced Pluripotent Stem Cells and Human Disease: Methods and Protocols serves as a vital guide that is valuable for not only experts but also novices in the stem cell field.

Stem cells have generated a lot of excitement among the researchers, clinicians and the public alike. Various types of stem cells are being evaluated for their regenerative potential. Marginal benefit resulting by transplanting autologus stem cells (deemed to be absolutely safe) in various clinical conditions has been proposed to be a growth factor effect rather than true regeneration. In contrast, various pre-clinical studies have been undertaken, using differentiated cells from embryonic stem cells or induced pluripotent stem cells have shown promise, functional improvement and no signs of teratoma formation. The scientists are not in a rush to reach the clinic but a handful of clinical studies have shown promise. This book is a collection of studies/reviews, beginning with an introduction to the pluripotent stem cells and covering various aspects like derivation, differentiation, ethics, etc., and hence would provide insight into the recent standing on the pluripotent stem cells biology. The chapters have been categorized into three sections, covering subjects ranging from the generation of pluripotent stem cells and various means of their derivation from embryonic as well as adult tissues, the mechanistic understanding of pluripotency and narrating the potential therapeutic implications of these in vitro generated cells in various diseases, in addition to the associated pros and cons in the same.

Induced Pluripotent Stem (iPS) cells are mature cells that have been genetically reprogrammed so that they return to their embryonic state. It is not yet known if iPS stem cells and embryonic stem cells differ significantly. Today many fundamental belief systems in biology are shifting towards accepting that mature body cells can be reverted to an embryonic state without the help of eggs or embryos. With their changed identities, iPS cells are then ready to serve as new tools for research in the fields of disease pathogenesis, drug discovery, oncology, and cell transplantation. One example of this would be using iPS from a patient’s mature cells to repair damaged tissue; it is thought that there would be very low incidence of rejection of the ‘new’ tissue in these cases. For the last four years, this therapeutic promise has been studied by hundreds of researchers worldwide in an effort to understand the ability of these cells to reverse their biological clocks.

A discussion of all the key issues in the use of human pluripotent stem cells for treating degenerative diseases or for replacing tissues lost from trauma. On the practical side, the topics range from the problems of deriving human embryonic stem cells and driving their differentiation along specific lineages, regulating their development into mature cells, and bringing stem cell therapy to clinical trials. Regulatory issues are addressed in discussions of the ethical debate surrounding the derivation of human embryonic stem cells and the current policies governing their use in the United States and abroad, including the rules and conditions regulating federal funding and questions of intellectual property.

Pluripotent stem cells have distinct characteristics: self-renewal and the potential to differentiate into various somatic cells. In recent years, substantial advances have been made from basic science to clinical applications. The vast amount knowledge available makes obtaining concise yet sufficient information difficult, hence the purpose of this book. In this book, embryonic stem cells, induced pluripotent stem cells, and mesenchymal stem cells are discussed. The book is divided into five sections: pluripotency, culture methods, toxicology, disease models, and regenerative medicine. The topics covered range from new concepts to current technologies. Readers are expected to gain useful information from expert contributors.

Brain diseases can have a large impact on patients and society, and treatment is often not available. A new approach in which somatic cells are reprogrammed into induced pluripotent cells (iPS cells) is a significant breakthrough for regenerative medicine. This promises patient-specific tissue for replacement therapies, as well as disease-specific cells for developmental modeling and drug treatment screening. However, this method faces issues of low reprogramming efficiency, and poorly defined criteria for determining the conversion of one cell type to another. Cells contain epigenetic “memories” of what they were that can affect reprogramming. This book discusses the various methods to reprogram cells, the control and determination of cell identity, the epigenetic models that have emerged and the application of iPS cell therapy for brain diseases, in particular Parkinson’s disease and Vanishing White Matter (VWM).​

Comprehensive coverage of the entire induced pluripotent stem cell basic work flow Pluripotent stem cells (PSC) can divide indefinitely, self-renew, and can differentiate to functionally reconstitute almost any cell in the normal developmental pathway, given the right conditions. This comprehensive book, which was developed from a training course, covers all of the PSCs (embryonic, embryonic germ, and embryonic carcinoma) and their functions. It demonstrates the feeder-dependent and feeder-free culture of hESC and hiPSC, which will be referred to in all protocols as PSCs. It also addresses the methods commonly used to determine pluripotency, as defined by self-renewal marker expression and differentiation potential. Human Pluripotent Stem Cells: A Practical Guide offers in-depth chapter coverage of introduction to stem cell, PSC culture, reprogramming, differentiation, PSC characterization, and more. It also includes four appendixes containing information on reagents, medias, and solutions; common antibodies; consumable and equipment; and logs and forms. Includes helpful tips and tricks that are normally omitted from regular research papers Features useful images to support the technical aspects and results visually as well as diagrammatic illustrations Presents specific sections (ie: reprogramming, differentiation) in a concise and easily digestible manner Written by experts with extensive experience in stem cell technologies Human Pluripotent Stem Cells: A Practical Guide is an ideal text for stem cell researchers, including principal investigators, and others in university and industry settings, and for new graduate students in PSC labs.