Human Pluripotent Stem Cells

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 BiologyTM 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.

This lavishly-illustrated, authoritative atlas explores the intricate art of culturing human pluripotent stem cells. Twelve chapters – containing more than 280 color illustrations – cover a variety of topics in pluripotent stem cell culturing including mouse and human fibroblasts, human embryonic stem cells and induced pluripotent stem cells, characteristic staining patterns, and abnormal cultures, among others. Atlas of Human Pluripotent Stem Cells in Culture is a comprehensive collection of illustrated techniques complemented by informative and educational captions examining what good quality cells look like and how they behave in various environments. Examples of perfect cultures are compared side-by-side to less-than-perfect and unacceptable examples of human embryonic and induced pluripotent stem cell colonies. This detailed and thorough atlas is an invaluable resource for researchers, teachers, and students who are interested in or working with stem cell culturing.

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 volume provides readers with a comprehensive collection of methods to guide them on how to generate, characterize, and use naïve human pluripotent stem cells (hPSCs). The chapters in this book cover topics such as three predominant routes to generate naïve hPSC lines; methods to differentiate naïve hPSCs into specialized cell types; and techniques to characterize naïve hPSCs using key molecular landmarks that benchmark and quality control the cell lines. 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. Cutting-edge and thorough, Human Naïve Pluripotent Stem Cells: Methods and Protocols is a valuable resource for novice and expert researchers who are looking to learn more or expand their research in this developing field.

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.

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 manual is a comprehensive compilation of "methods that work" for deriving, characterizing, and differentiating hPSCs, written by the researchers who developed and tested the methods and use them every day in their laboratories. The manual is much more than a collection of recipes; it is intended to spark the interest of scientists in areas of stem cell biology that they may not have considered to be important to their work. The second edition of the Human Stem Cell Manual is an extraordinary laboratory guide for both experienced stem cell researchers and those just beginning to use stem cells in their work. Offers a comprehensive guide for medical and biology researchers who want to use stem cells for basic research, disease modeling, drug development, and cell therapy applications. Provides a cohesive global view of the current state of stem cell research, with chapters written by pioneering stem cell researchers in Asia, Europe, and North America. Includes new chapters devoted to recently developed methods, such as iPSC technology, written by the scientists who made these breakthroughs.

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.