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This book explores the vital importance of T-cell differentiation in areas as wide-ranging as pathological analysis, drug development, and cell therapy of human T-cells. Focusing on human embryonic stem cells and human induced pluripotent stem cells, the chapters explore a variety of in vitro T-cell differentiation protocols as well as useful techniques to develop and evaluate cellular medicines. 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. Authoritative and practical, In Vitro Differentiation of T-Cells: Methods and Protocols serves as an ideal guide for researchers seeking to differentiate T-cells from pluripotent stem cells in order to achieve any number of significant goals.
This volume provides protocols to successfully apply cutting-edge technologies to characterize the biology of T cells at an unprecedented level of complexity. Chapters guide readers through flow cytometry and fluorescence-activated cell sorting, the behaviour of single T cells after adoptive cell transfer (ACT), single cell gene expression by multiplex PCR, lentiviral transduction approaches, protocols to derive large numbers of early-differentiated memory T cells by using dedicated cytokines cocktails, approaches to measure telomerase activity in terminally differentiated T cells, and approaches to define Treg cells at the phenotypic and functional level. The final part of the book is dedicated to the analysis of the differentiation and effector functions of innate T cells, namely the well-known γ/δ T cells, and the recently identified CD8] mucosal associated invariant T (MAIT) 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. Authoritative and cutting-edge, T-Cell Differentiation: Methods and Protocols aims to provide protocols that are fundamental to monitor the T cell compartment at the level of single cells in pathological and immunotherapy conditions.
Current Protocols in Immunology is a three-volume looseleaf manual that provides comprehensive coverage of immunological methods from classic to the most cutting edge, including antibody detection and preparation, assays for functional activities of mouse and human cells involved in immune responses, assays for cytokines and their receptors, isolation and analysis of proteins and peptides, biochemistry of cell activation, molecular immunology, and animal models of autoimmune and inflammatory diseases. Carefully edited, step-by-step protocols replete with material lists, expert commentaries, and safety and troubleshooting tips ensure that you can duplicate the experimental results in your own laboratory. Bimonthly updates, which are filed into the looseleaf, keep the set current with the latest developments in immunology methods. The initial purchase includes one year of updates and then subscribers may renew their annual subscriptions. Current Protocols publishes a family of laboratory manuals for bioscientists, including Molecular Biology, Human Genetics, Protein Science, Cytometry, Cell Biology, Neuroscience, Pharmacology, and Toxicology.
Aging is an almost universal process within biological systems, one which leads to a decline in functional capacity, disease onset, and eventually death. There has been much interest in recent years to elucidate the molecular mec- nisms that underlie the aging process. Many theories have been proposed since the last century that aim to explain the causes of aging. There is no one theory that completely satisfies the phenotype of aging, but genetics and environm- tal factors play an important role in the etiology of age-related pathologies and the aging process. However, there is still much to be learned about the aging process which has been termed one of the last great frontiers in biology. De- graphic changes worldwide are leading to increased average life expectancies within our populations. These changes in population characteristics will impact upon the economies of the supporting society, with increasing healthcare and infrastructural costs arising from the prevalence of age-related pathologies and other physical disabilities associated with advancing years. Many researchers worldwide are working in the attempt to identify key cellular processes through which it might one day be possible to slow down the aging process and thus increase the health span of humans. Numerous research projects—from the cellular through to tissue, organ, and whole organism studies—are currently underway to investigate the mul- factorial aging process.
This volume presents a broad selection of cutting-edge methods and tools that will enable the reader to investigate the multi-faceted manifestations of inflammation. Inflammation: Methods and Protocols is divided into four sections: the first three sections describe protocols investigating immune-mediated inflammatory disease models affecting barrier organs to the environment; the skin, the lung, and the intestinal and oral mucosa. The fourth section illustrates inflammatory disease models of the brain, joints, and vasculature. 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 notes on troubleshooting and avoiding known pitfalls. Authoritative and practical, Inflammation: Methods and Protocols aims to inspire the experienced investigator and the young experimenter alike to disentangle the fascinating process of inflammation.
This volume provides comprehensive methods from expert scientists working in the Chimeric Antigen Receptor T Cell (CAR-T Cell) field. Chapters guide readers through the state-of-art of CAR-T cell technology, CAR design and vector production, CAR-T cell generation and manufacturing, CAR-T cell characterization, and quality control. 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 cutting-edge, Chimeric Antigen Receptor T Cells: Development and Production aims to be useful in the production of CAR-T cells, especially for therapeutic purposes.
This second edition volume provides new and updated chapters detailing simple and accessible experiment protocols to explore thymus biology. Chapters are divided into three parts presenting short reviews, analysis strategies, protocols for cell preparation, flow cytometry analyses, Innate Lymphoid Cells (ILC), mouse T-cell development, antigen receptor-less cousins of T cells, bone marrow chimeras, thymic stroma, and multiple aspects of thymocyte biology. 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 cutting-edge, T-Cell Development: Methods and Protocols, Second Edition aims to be a useful practical guide to help readers overcome obstacles associated with experimental approaches of T-cell development.
This volume provides methods and models for investigating the immunosuppressive subset of CD4+ T-cells, regulatory T-cells (Treg). Chapters guide readers through, basic protocols for analyzing Treg in mice and humans, single-cell analysis for analyzing Treg, and methods for the analysis of Treg in clinical application. Written in the format of the highly successful Methods in Molecular Biology series, each chapter includes an introduction to the topic, lists necessary materials and reagents, includes tips on troubleshooting and known pitfalls, and step-by-step, readily reproducible protocols. Authoritative and cutting-edge, Regulatory T-Cells: Methods and Protocols aims to be a useful and practical guide to new researchers and experts looking to expand their knowledge.
At some point in their careers, virtually every scientist and technician, as well as many medical professionals, regardless of their area of specialization have a need to utilize cell culture systems. Updating and significantly expanding upon the previous editions, Basic Cell Culture Protocols, Fourth Edition provides the novice cell culturist with sufficient information to perform the basic techniques, to ensure the health and identity of their cell lines, and to be able to isolate and culture specialized primary cell types. The intent of this extensive volume is to generate a valuable resource containing clear methodologies pertinent to current areas of investigation, rather than attempting to educate cell culturists on specific cell types or organ systems. Written in the highly successful Methods in Molecular BiologyTM, 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, Basic Cell Culture Protocols, Fourth Edition compiles the essential techniques needed to approach this vital laboratory activity with full success.
This volume provides current and new advanced methods and protocols to study T cells. Chapters guide readers through T cell diversity using mass cytometry, analyzing T cells from single cell level, CRISPR/Cas9 techniques to study the T cell activation, techniques to study subsets of Tcell’s, procedures to study artificial antigen presentosomes for T cell activation, techniques to study the T cell development, two-photon microscopy, and MAIT 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. Authoritative and cutting-edge, T-Cell Receptor Signaling: Methods and Protocols aims to provide a wide range of approaches and be an invaluable resource for present and future generations of T cell researchers.