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Full Description
This detailed volume explores the use of Sertoli cells as models in male reproductive biology or as supporters for other cell types, and it answers the need for standardized protocols for their in vitro culture. Prepared for non-specialists as well as experienced researchers that may be interested in a multidisciplinary approach to study these cells, the book examines these remarkable cells that present some unique features that go far beyond the physical and nutritional support of spermatogenesis. Written for 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, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.
Authoritative and practical, Sertoli Cells: Methods and Protocols serves as a valuable resource for experienced researchers in reproductive biology, particularly andrologists, but also for future investigators and young students starting to study these extraordinary cells.
Contents
Establishment of Primary Culture of Sertoli Cells.- Evaluation of the Purity of Sertoli Cell Primary Cultures.- Preparation of Testicular Samples for Histology and Immunohistochemistry.- Rabbit Sertoli Cells: Immunohistochemical Profile from Neonatal to Adult Age.- Identification of Proliferative and Apoptotic Sertoli Cells Using Fluorescence and Confocal Microscopy.- Sertoli Cells Preparation for Co-Immunoprecipitation.- Profiling Signaling Proteins in Sertoli Cells by Co-Immunoprecipitation.- Phagocytosis by Sertoli Cells: Analysis of Main Phagocytosis Steps by Confocal and Electron Microscopy.- A Method for In Vivo Induction and Ultrastructural Detection of Mitophagy in Sertoli Cells.- Assessing Autophagy in Sertoli Cells.- Molecular Mechanisms and Signaling Pathways Involved in the Nutritional Support of Spermatogenesis by Sertoli Cells.- Assessing Sertoli Cells Metabolic Activity.- Proteome Profiling of Sertoli Cells Using a GeLC-MS/MS Strategy.- Gene Silencing of Human Sertoli Cells Utilizing Small Interfering RNAs.- Testicular Cell Selective Ablation Using Diphtheria Toxin Receptor Transgenic Mice.- Regulation of Blood-Testis Barrier (BTB) Dynamics, Role of the Actin, and Microtubule-Based Cytoskeletons.- Monitoring the Integrity of the Blood-Testis Barrier (BTB): An In Vivo Assay.- Computational Methods Involved in Evaluating the Toxicity of the Reproductive Roxicants in Sertoli Cell.- A Stopped-Flow Light Scattering Methodology for Assessing the Osmotic Water Permeability of Whole Sertoli Cells.- Cryopreservation of Human Testicular Tissue by Isopropyl-Controlled Slow Freezing.
