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Scientists investigating germ cells have, over the past 15 years, originated discoveries and innovations that give us valuable insights into the mechanisms that regulate not just stem cell function, but human development in its widest sense. With contributions from some of the leading researchers in the field, Male Germline Stem Cells: Developmental and Regenerative Potential assesses the implications of these discoveries for understanding the fundamental biology of germline stem cells as well as their potential for human stem cell-based therapies. This monograph covers many of the fundamental issues now being explored by today’s generation of stem cell researchers, including the field’s potential for regenerative medicine. Ranging from an assessment of the pluripotency of primordial germ cells and their possible applications in treating testicular cancer, to the recovery of once-mordant fertilization-competent sperm, this volume has it all. It is a reference point for any scientist involved in related research as well as being a timely summation of what could prove to be a hugely exciting and very fruitful area of inquiry.
In this comprehensive and cutting-edge book, leading experts explore the parameters that define germline stem cells and the mechanisms that regulate the cell behavior in order to better isolate, characterize and maintain them. The volume begins by providing protocols for germline stem cell identification and regulation in model organisms, and concludes with detailed chapters covering current techniques involving in vitro culture and the applications of the cells.
Drosophila melanogaster: Practical Uses in Cell and Molecular Biology is a compendium of mostly short technical chapters designed to provide state-of-the art methods to the broad community of cell biologists, and to put molecular and cell biological studies of flies into perspective. The book makes the baroque aspects of genetic nomenclature and procedure accessible to cell biologists. It also contains a wealth of technical information for beginning or advanced Drosophila workers. Chapters, written within a year of publication, make this topical volume a valuable laboratory guide today and an excellent general reference for the future. Key Features* Collection of ready-to-use, state-of-the art methods for modern cell biological and related research using Drosophila melanogaster* Accessible to both experienced Drosophila researchers and to others who wish to join in at the cutting edge of this system * Drosophila offers an easily managed life cycle, inexpensive lifestyle, extraordinarily manipulable molecular and classical genetics, now combined with powerful new cell biology techniques * Introduction and overview sections orient the user to the Drosophila literature and lore * Six full-color plates and over 100 figures and tables enhance the understanding of these cell biology techniques
This volume explores the latest techniques used in the field of mammalian germ cell biology to study key experimental approaches and perspectives for critical analysis of spermatogonial subsets including the rare spermatogonial stem cells (SSCs) pool. 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 common pitfalls. Cutting-edge and comprehensive, Spermatogonial Stem Cells: Methods and Protocols is a valuable resource for both new and experiences investigators looking to expand their knowledge in this developing field.
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.
Eggs of all animals contain mRNAs and proteins that are supplied to or deposited in the egg as it develops during oogenesis. These maternal gene products regulate all aspects of oocyte development, and an embryo fully relies on these maternal gene products for all aspects of its early development, including fertilization, transitions between meiotic and mitotic cell cycles, and activation of its own genome. Given the diverse processes required to produce a developmentally competent egg and embryo, it is not surprising that maternal gene products are not only essential for normal embryonic development but also for fertility. This review provides an overview of fundamental aspects of oocyte and early embryonic development and the interference and genetic approaches that have provided access to maternally regulated aspects of vertebrate development. Some of the pathways and molecules highlighted in this review, in particular, Bmps, Wnts, small GTPases, cytoskeletal components, and cell cycle regulators, are well known and are essential regulators of multiple aspects of animal development, including oogenesis, early embryogenesis, organogenesis, and reproductive fitness of the adult animal. Specific examples of developmental processes under maternal control and the essential proteins will be explored in each chapter, and where known conserved aspects or divergent roles for these maternal regulators of early vertebrate development will be discussed throughout this review. Table of Contents: Introduction / Oogenesis: From Germline Stem Cells to Germline Cysts / Oocyte Polarity and the Embryonic Axes: The Balbiani Body, an Ancient Oocyte Asymmetry / Preparing Developmentally Competent Eggs / Egg Activation / Blocking Polyspermy / Cleavage/ Mitosis: Going Multicellular / Maternal-Zygotic Transition / Reprogramming: Epigenetic Modifications and Zygotic Genome Activation / Dorsal-Ventral Axis Formation before Zygotic Genome Activation in Zebrafish and Frogs / Maternal TGF-β and the Dorsal-Ventral Embryonic Axis / Maternal Control After Zygotic Genome Activation / Compensation by Stable Maternal Proteins / Maternal Contributions to Germline Establishment or Maintenance / Perspective / Acknowledgments / References
Stem cell science has the potential to impact human reproductive medicine significantly - cutting edge technologies allow the production and regeneration of viable gametes from human stem cells offering potential to preciously infertile patients. Written by leading experts in the field Stem Cells in Reproductive Medicine brings together chapters on the genetics and epigenetics of both the male and female gametes as well as advice on the production and regeneration of gene cells in men and women, trophoblasts and endometrium from human embryonic and adult stem cells. Although focussing mainly on the practical elements of the use of stem cells in reproductive medicine, the book also contains a section on new developments in stem cell research. The book is essential reading for reproductive medicine clinicians, gynecologists and embryologists who want to keep abreast of practical developments in this rapidly developing field.
Telomerase is required for maintaining the male germline in worms, fish and mice. Underlying the continuous production of sperm in these species are poorly characterized germline stem and progenitors. Telomere attrition is strictly avoided during sperm production, thus avoiding the inter-generational transmission of ageing phenotypes. The work described in this dissertation aims to answer fundamental questions regarding the behavior of germline stem cells and the control of telomerase within those cells. Firstly, we describe the generation of telomerase reverse transcriptase reporter mice, allowing the prospective isolation of germ cells with discrete telomerase levels, and uncovering the basis principles of telomerase regulation in vivo. We discover that high telomerase levels are a hallmark of the male germline stem cell lineage, regardless of proliferation status, and that telomere attrition depletes the stem cell pool. Secondly, we exploit the pattern of telomerase expression in the adult testis to understand germline stem cell heterogeneity. We fractionate purified spermatogonia into GFRa1+ and GFRa1- populations, enabling functional studies by transplantation and molecular characterization by RNA-Seq. Although GFRa1+ cells were thought to represent the predominant self-renewing population by lineage tracing, GFRa1+ and GFRa1- cells exhibit comparably high stem cell activity by transplantation. The transition from GFRa1+ to GFRa1- is accompanied by a reduction in GDNF and FGF signaling and a marked decrease in proliferation. Upon transplantation, GFRa1- cells replenish the GFRa1+ population, revealing that GFRa1- cells are in a poised state, retaining stem cell potential and the ability to be reprogrammed by a vacant niche. In the final part of this dissertation, we again exploit TERT-reporter mice to determine the transcriptomes of defined spermatogonial subtypes. These datasets established a pipeline for the rapid discovery and validation of new markers for undifferentiated spermatogonia (MSI2 and SDC4), differentiated spermatogonia (ALCAM), GFRa1- cells (OCT4-delta-PE-GFP), and finally GFRa1+ cells (MCAM).
This second edition provides updated and new chapters on selected genetic, molecular, biochemical, and cell biological techniques. Chapter’s guide readers through methods and principles on primordial germ cells and germline stem cells, however many of these principles can be applied to different types of adult 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. Authoritative and cutting-edge, Germline Stem Cells: Second Edition aims to present the new findings and techniques that have developed since the first edition.