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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.
The volume provides a comprehensive overview of methods that have been developed for the study of Tfh and Tfr biology in mice, and in different human diseases. The chapters detail methods to investigate the function of Tfh cells in vivo and in vitro, including the use of mathematical models to study GC dynamics. 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-Follicular Helper Cells: Methods and Protocols aims to be a useful practical guide to researchers to help further their study in this field.
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.
This volume presents the latest collection of immunophenotypic techniques and applications used in research and clinical settings. Chapters in this book cover topics such as constructions of high dimensions fluorescence and mass cytometry panels; fluorescence barcoding; using dried or lyophilized reagents; and immunophenotypic examples of specific cell types. The book concludes with a discussion on the critical roles of quality control and immunophenotyping in the clinical environment. 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 comprehensive, Immunophenotyping: Methods and Protocols is a valuable resource for any researchers, clinician, or scientist interested in learning more about this evolving field.
B-lymphocyte development and function remains an exciting area of research for those interested in the physiology and pathology of the immune system in higher animals. While recent advances in genetics and cellular and molecular biology have provided a large spectrum of powerful new experimental tools in this field, it is both time consuming and often very difficult for a student or just any bench-side worker to identify a reliable experimental protocol in the ocean of the literature. The aim of B Cell Protocols is to provide a collection of diverse protocols ranging from the latest inventions and applications to some classic, but still frequently used methods in B-cell biology. The authors of the various chapters are all highly qualified scientists who are either the inventors or expert users of these methods. Their extensive experience in mastering a particular method provides not only the step-by-step details of a reproducible protocol, but also useful troubleshooting tips that readers will appreciate in their daily work. We hope that this book will be helpful for both beginning and experienced researchers in the field in designing or modifying an experimental approach, and exploring a biological question from multiple angles.
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.