This book pulls together the full range of cell culture, biochemical, microscopic, and genetic techniques to study the early mammalian embryo. Until now, there has never been such a comprehensive compendium, though there have been more focused books of protocol, such as "Manipulating the Mouse Embryo", from Cold Spring Harbor. This book is intended to appeal to all constituencies, from basic experimental science to clinical and animal science applications.
Preface; Acknowledgements; List of Contributors; Abbreviations; 1. Follicle development in vitro; 2. Preparation of gametes, in vitro maturation, in vitro fertilization and embryo recovery; 3. Culture of the mammalian preimplantation embryo; 4. Assessment of embryo development and viability; 5. Micromanipulation : ICSI; 6. Micromanipulation : Biopsy; 7. Analysis of intracellular ions: pH and calcium; 8. Nutrient uptake and metabolite production and enzyme activity and regulation; 9. Metabolic Pathway; 10. Confocal imaging of structural molecules in mammalian gametes; 11. Fluorescent in situ hybridization; 12. Assessment of ploidy, telomere length and telomerase activity in oocytes and embryos; 13. Microscale RNA isolation from mammalian embryos; 14. Relative mRNA transcript abundance in early embryos by Reverse Transcription-Polymerase Chain Reaction (RT-PCR); 15. Characterization of novel genes during early development by application of differential display RT-PCR; 16. Two-dimensional protein gel database analysis of embryos, oocytes, and oocyte-associated granulosa cells; 17. Antisense disruption of gene expression in the preimplantation embryo; 18. Gene subtraction and analysis; 19. Analysis of apoptosis in the preimplantation embryo; 20. Cytoplasmic Signaling and Cell Cycle Control in the Mouse Egg and Embryo; 21. Human Embryonic Stem; 22. Spermatogonial Transplantion; 23. Somatic Cell Nuclear Transfer; 24. Cryobiology; slow freezing vitrification of embryos