Nuclear reprogramming : methods and protocols / edited by Steve Pells.
Contributor(s): Pells, SteveMaterial type: TextSeries: Methods in molecular biology (Clifton, N.J.): v. 325.Publisher: Totowa, N.J. : Humana Press, ©2006Description: 1 online resource (xiv, 333 pages) : illustrations (some color)Content type: text Media type: computer Carrier type: online resourceISBN: 1588293793; 1597450057; 9781597450058; 9781588293794; 1280358602; 9781280358609Subject(s): Cell nuclei -- Transplantation -- Laboratory methods | 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 | Noyau (Cellule) -- Transplantation -- Manuels de laboratoire | SCIENCE -- Biotechnology | Cell nuclei -- Transplantation | Biology | Health & Biological Sciences | Cytology | GeneticsGenre/Form: Electronic books. | Electronic books. Additional physical formats: Print version:: Nuclear reprogramming.DDC classification: 660.6/5 LOC classification: QH442.2 | .N84 2006Online resources: Click here to access online
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Includes bibliographical references and index.
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.
Print version record.
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.