Title Nuclear reprogramming : methods and protocols / edited by Steve Pells

Published Totowa, N.J. : Humana Press, ©2006

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Description 1 online resource (xiv, 333 pages) : illustrations (some color) Series Methods in molecular biology, 1064-3745 ; 325 Methods in molecular biology (Clifton, N.J.) ; v. 325 Contents Nuclear transplantation in xenopus / John B. Gurdon -- Nuclear transfer in sheep / William A. Ritchie -- Protocols for nuclear transfer in mice / Shaorong Gao -- Isolation of stromal stem cells from human adipose tissue / Andrew C. Boquest [and others] -- Nuclear reprogramming by cell fusion / Robert H. Broyles [and others] -- Polyethylene glycol-mediated cell fusion / Jian Yang and Ming Hong Shen -- Epigenetic reprogramming of somatic genomes by electrofusion with embryonic stem cells / Masako Tada and Takashi Tada -- Quantification of cell fusion by flow cytometry / Stephen Sullivan [and others] -- Modulation of cell fate using nuclear and cytoplasmic extracts / Anne-Mari Håkelien, Kristine G. Gaustad, and Philippe Collas -- Transgenic systems in nuclear reprogramming / Megan Munsie, Peter Mountford, and Jennifer Nichols -- Using immunofluorescence to observe methylation changes in mammalian preimplantation embryos / Fátima Santos and Wendy Dean -- Observing s-phase dynamic of histone modifications with fluorescently labeled antibodies / Rong Wu, Anna V. Terry, and David M. Gilbert -- Quantitative analysis of telomerase activity and telomere length in domestic animal clones / Dean H. Betts [and others] Pluripotency: capacity for in vitro differentiation of undifferentiated embryonic stem cells / Cornelia Wiese [and others] -- Staining embryonic stem cells using monoclonal antibodies to stage-specific embryonic antigens / Bruce A. Fenderson [and others] -- Analysis of the nucleolar compartment of the nucleus as an indicator of nuclear reprogramming after nuclear transfer / Jacques-E. Fléchon -- Methylation-sensitive polymerase chain reaction / Hannah R. Moore, Richard R. Meehan, and Lorraine E. Young -- Analysis of DNA methylation profiles in preimplantation embryos using bisulfite mutagenesis / Yong-Mahn Han, Seok-Ho Kim, and Yong-Kook Kang -- Chromatin immunoprecipitation assay for mammalian tissues / Fiona B. Turner, Wang L. Cheung, and Peter Cheung -- Histone modifications and transcription factor binding on chromatin: CHIP/PCR assays / Jaejoon Won and Tae Kook Kim -- In vivo genomic footprinting using LM-PCR methods / Hiromi Tagoh, Peter N. Cockerill, and Constanze Bonifer -- Analyzing histone modification using crosslinked chromatin treated with micrococcal nuclease / Pascal Lefevre and Constanze Bonifer Summary Although nuclear reprogramming-the cellular process underlying cloning and the artificial generation of stem cells from somatic cells-may be considered a "black box" whose interior events are either completely unknown or at least imperfectly characterized, tantalizing clues about its operation are beginning to emerge. In Nuclear Reprogramming: Methods and Protocols, hands-on laboratory scientists describe readily reproducible methods for actually performing nuclear reprogramming by nuclear transfer in several different species, by fusion induced both chemically and electrically, and by the in vivo treatment of cells with cell extracts. Isolation of an adult stem cell type is also described. In addition, several methods of monitoring nuclear reprogramming are presented, including the use of transgenic markers, activation of telomerase as an ES-specific marker, light and electron microscopic observation of structural changes in the nucleus, and the verification of surface marker expression and differentiation potential of stem cells. Biochemical methods are provided for the examination of chromatin protein modifications, nucleosomal footprinting, transcription factor binding, and the study of DNA methylation changes both at the specific locus level and at the level of the whole nucleus. The protocols follow the successful Methods in Molecular Biology series format, each offering step-by-step laboratory instructions, an introduction outlining the principles behind the technique, lists of the necessary equipment and reagents, and tips on troubleshooting and avoiding known pitfalls. Wide-ranging and highly practical, Nuclear Reprogramming: Methods and Protocols significantly expands the repertoire of methods available to perform, study, and understand nuclear programming, not only for cloning, but also for reprogramming somatic cells to stem cells Bibliography Includes bibliographical references and index Notes English Print version record Subject Cell nuclei -- Transplantation -- Laboratory methods Cell organelles. Cell physiology. Cytology -- Technique. Surgery, Operative. Life sciences. Physical sciences. Medical laboratory technology. Cells. Cytoplasm. Anatomy. Cell nuclei. Cell hybridization. Transplantation of organs, tissues, etc. Methodology. Physiology. Cell nuclei -- Transplanting -- Laboratory manuals Cell Nucleus -- physiology Cell Fusion -- methods Transplantation -- methods Intracellular Space Organelles Investigative Techniques Cell Physiological Phenomena Cytological Techniques Surgical Procedures, Operative Biological Science Disciplines Natural Science Disciplines Analytical, Diagnostic and Therapeutic Techniques and Equipment Cytoplasmic Structures Cellular Structures Clinical Laboratory Techniques Phenomena and Processes Cells Cytoplasm Disciplines and Occupations Anatomy Cell Nucleus Cell Fusion Transplantation Methods Physiology biological sciences. physical sciences. anatomy. methodology. physiology. SCIENCE -- Biotechnology. Transplantation of organs, tissues, etc. Surgery, Operative Physiology Physical sciences Methodology Medical laboratory technology Life sciences Cytoplasm Cytology -- Technique Cells Cell physiology Cell organelles Cell nuclei Cell hybridization Anatomy Cell nuclei -- Transplantation Genre/Form Laboratory manuals Laboratory manuals. Manuels de laboratoire. Form Electronic book Author Pells, Steve ISBN 1588293793 1597450057 9781597450058 9781588293794 1280358602 9781280358609 9786610358601 6610358605

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