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Full Description
This detailed volume examines techniques for better understanding mammalian preimplantation embryo development, the development and contribution of gametes, and how to manipulate such reproductive cells in the laboratory. Written by experts in their field, the book covers gamete preparation, in vitro embryo culture and cryopreservation, physiological and molecular methods for embryo assessment, and emerging methods for modeling key developmental processes encompassing gamete and embryo manipulation through implantation. As part of 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 and readily reproducible laboratory protocols, and tips for troubleshooting and avoiding known pitfalls.
Authoritative and practical, The Preimplantation Embryo: Methods and Protocols serves as a comprehensive and practical toolbox and will be of immense value to scientists working on the fascinating and dynamic preimplantation embryo and the gametes from which it originates.
Contents
In Vitro Development of Primordial Follicles from Human Ovarian Tissue.- Capacitation In Vitro Maturation (CAPA-IVM) of Human Oocytes.- Sperm Preparation in ART: Traditional Methods and Emerging Technologies.- Sperm Stratification by Combined Flow Cytometry-Based Determination of Sperm Samples for DNA Fragmentation and Membrane Integrity.- Sperm Proteomics and Phosphoproteomics: High-Throughput Methods Tailored to Gain Insight into Sperm Cell Signaling Pathways.- Conventional In Vitro Fertilization and Intracytoplasmic Sperm Injection.- Culture of the Human Embryo.- Oocyte and Embryo Vitrification.- Microwave-Assisted Dehydration, Non-Cryogenic Storage, and Rehydration of Spermatozoa and Germinal Vesicles.- Non-Invasive Assessment of Embryo Physiology.- Evaluating Preimplantation Embryo Metabolism Through Imaging of Cellular Autofluorescence.- Imaging Spatiotemporal RNA and Protein Dynamics in Mouse Embryos.- Embryo Biopsy.- Non-Invasive Preimplantation Genetic Testing (niPGT).- Analysis of Relative Telomere Length and mtDNA Content in Individual Oocytes and Embryos by Quantitative Real-Time PCR (qPCR).- Quantitative Analysis of Embryo Gene Expression by Real-Time PCR.- Analysis of mtDNA Methylation.- Single-Cell and Single-Embryo RNA Sequencing.- Immunofluorescence Analysis of Epigenetic Markers in Oocytes and Preimplantation Embryos.- Purification and Characterization of Extracellular Vesicles from Embryo-Conditioned Medium.- Modeling Early Human Embryo Implantation Using Human Endometrial Organoids and Embryo Surrogate Co-Cultures for MicroRNA Transfection.- Targeted Degradation of Mouse Embryo Proteins Using Trim-Away.- Somatic Cell Nuclear Transfer in Sheep and Approaches to Improve Efficiency.- Clustered Regularly Interspaced Palindromic Repeats (CRISPR) Applied to Gametes and Embryos.- Pronuclear Transfer and Maternal Spindle Transfer in the Mouse.
