Stem Cell Protocols

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.

Now in two volumes, this completely updated and expanded edition of Embryonic Stem Cells: Methods and Protocols provides a diverse collection of readily reproducible cellular and molecular protocols for the manipulation of nonhuman embryonic stem cells. Volume one, Embryonic Stem Cell Protocols: Isolation and Characterization, Second Edition, provides a diverse collection of readily reproducible cellular and molecular protocols for the isolation, maintenance, and characterization of embryonic stem cells. The second volume, Embryonic Stem Cell Protocols: Differentiation Models, Second Edition, covers state-of-the-art methods for deriving many types of differentiating cells from ES cells. Together, the two volumes illuminate for both novices and experts our current understanding of the biology of embryonic stem cells and their utility in normal tissue homeostasis and regenerative medicine applications.

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.

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 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.

Since 1998, the volume of research being conducted using human embryonic stem (hES) cells has expanded primarily using private funds because of restrictions on the use of federal funds for such research. Given limited federal involvement, privately funded hES cell research has thus far been carried out under a patchwork of existing regulations, many of which were not designed with this research specifically in mind. In addition, hES cell research touches on many ethical, legal, scientific, and policy issues that are of concern to the public. This report provides guidelines for the conduct of hES cell research to address both ethical and scientific concerns. The guidelines are intended to enhance the integrity of privately funded hES cell research by encouraging responsible practices in the conduct of that research.

This revised edition provides up-to-date protocols developed in the HSC field. A team of leading researchers supply this volume with in-depth, readily reproducible methods for effective characterization of HSC and their developmental potential. The book provides detailed flow cytometry protocols for thorough analysis of enriched HSC populations, and offers a variety of transplantation approaches to measure HSC function in vivo. This is a much needed technical resource in the critically important field of stem cell investigation.