Germline Stem Cells Methods and Protocols

This third edition volume expands on the previous editions with new discussion on the latest techniques and model systems used to study germline stem cells. The chapters in this book cover topics such as analysis of the C. elegans germline stem cell pool; live imaging of the Drosophila testis stem cell niche; standardization of single-cell RNA-sequencing analysis workflow to study Drosophila ovary; 3D analysis of the intact mouse ovary; and derivation and primordial germ cell induction of intermediate pluripotent stem cells. Written in the highly successful Methods in Molecular Biology series format, 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.

Cutting-edge and practical, Germline Stem Cells: Methods and Protocols, Third Edition is a valuable resource for all researchers in the germ stem cell community interested in learning more about this developing field.

 Analysis of the C. elegans Germline Stem Cell Pool.- Analysis of C. elegans Germline Small RNA Pathways.- Lineage Tracing and Single Cell RNA-seq in C. elegans to Analyze Transgenerational Epigenetic Phenotypes Inherited from Germ Cells.- Methods to Analyze Nutritional and Inter-Organ Control of Drosophila Ovarian Germline Stem Cells.- Targeting Endogenous loci that Function in the Drosophila Germline Stem Cells.- Live Imaging of the Drosophila Testis Stem Cell Niche.- Isolation of Non-Tumorous Germline and Somatic Cells from Drosophila Testes.- Spermatogonial Dedifferentiation into Germline Stem Cells in Drosophila Testes.- Standardization of Single-Cell RNA-Sequencing Analysis Workflow to Study Drosophila Ovary.- Identification of Germline Stem Cells in Zebrafish.- Characterization of the Postnatal Naked Mole-Rat Ovary: From Primordial Germ Cells to Meiotic Prophase I Oocytes.- A Roadmap for Three-Dimensional Analysisof the Intact Mouse Ovary.- Isolation and Culture Techniques for Fetal Mouse Germ Cells.- Rattus norvegicus Spermatogenesis Colony Forming Assays.- Maintenance of Human Primordial Germ Cell-Like Cells in a Long-Term Culture System.- Derivation and Primordial Germ Cell Induction of Intermediate Pluripotent Stem Cells.