Nuclear Reprogramming

1. Front Matter

   Pages i-xiv

   PDF

2. Nuclear Transplantation in Xenopus

   • John B. Gurdon

   Pages 1-9

3. Nuclear Transfer in Sheep

   • William A. Ritchie

   Pages 11-23

4. Protocols for Nuclear Transfer in Mice

   • Shaorong Gao

   Pages 25-33

5. Isolation of Stromal Stem Cells From Human Adipose Tissue

   • Andrew C. Boquest, Aboulghassem Shahdadfar, Jan E. Brinchmann, Philippe Collas

   Pages 35-46

6. Nuclear Reprogramming by Cell Fusion

   • Robert H. Broyles, Austin C. Roth, Mairead Todd, Visar Belegu

   Pages 47-57

7. Polyethylene Glycol-Mediated Cell Fusion

   • Jian Yang, Ming Hong Shen

   Pages 59-66

8. Epigenetic Reprogramming of Somatic Genomes by Electrofusion With Embryonic Stem Cells

   • Masako Tada, Takashi Tada

   Pages 67-79

9. Quantification of Cell Fusion by Flow Cytometry

   • Stephen Sullivan, Martin Waterfall, Ed J. Gallagher, Jim McWhir, Steve Pells

   Pages 81-97

10. Modulation of Cell Fate Using Nuclear and Cytoplasmic Extracts

    • Anne-Mari Håkelien, Kristine G. Gaustad, Philippe Collas

    Pages 99-114

11. Transgenic Systems in Nuclear Reprogramming

    • Megan Munsie, Peter Mountford, Jennifer Nichols

    Pages 115-128

12. Using Immunofluorescence to Observe Methylation Changes in Mammalian Preimplantation Embryos

    • Fátima Santos, Wendy Dean

    Pages 129-138

13. Observing S-Phase Dynamics of Histone Modifications With Fluorescently Labeled Antibodies

    • Rong Wu, Anna V. Terry, David M. Gilbert

    Pages 139-148

14. Quantitative Analysis of Telomerase Activity and Telomere Length in Domestic Animal Clones

    • Dean H. Betts, Steven Perrault, Lea Harrington, W. Allan King

    Pages 149-180

15. Pluripotency: Capacity for In Vitro Differentiation of Undifferentiated Embryonic Stem Cells

    • Cornelia Wiese, Gabriela Kania, Alexandra Rolletschek, Przemyslaw Blyszczuk, Anna M. Wobus

    Pages 181-205

16. Staining Embryonic Stem Cells Using Monoclonal Antibodies to Stage-Specific Embryonic Antigens

    • Bruce A. Fenderson, Maria P. De Miguel, April D. Pyle, Peter J. Donovan

    Pages 207-224

17. Analysis of the Nucleolar Compartment of the Nucleus as an Indicator of Nuclear Reprogramming After Nuclear Transfer

    • Jacques-E. Fléchon

    Pages 225-238

18. Methylation-Sensitive Polymerase Chain Reaction

    • Hannah R. Moore, Richard R. Meehan, Lorraine E. Young

    Pages 239-249

19. Analysis of DNA Methylation Profiles in Preimplantation Embryos Using Bisulfite Mutagenesis

    • Yong-Mahn Han, Seok-Ho Kim, Yong-Kook Kang

    Pages 251-260

20. Chromatin Immunoprecipitation Assay for Mammalian Tissues

    • Fiona B. Turner, Wang L. Cheung, Peter Cheung

    Pages 261-272

A wide-ranging collection of readily reproducible methods for performing nuclear reprogramming by nuclear transfer in several different species, by fusion through both chemical treatment and electrically shocking cells, and by in vivo treatment of cells with cell extracts. Several methods of monitoring nuclear reprogramming are also 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 verification of surface marker expression and the 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.

• Activation
• DNA
• Ethylen
• Monoclonal Antibodies
• PCR
• Telomere
• Vivo
• transcription

• Department of Gene Function and Development, Roslin Institute, Roslin

  Steve Pells

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