Germline Stem Cells, 2008 Methods in Molecular Biology Series, Vol. 450

Stem cells are responsible for maintaining tissue homeostasis. They possess unlimited self-renewal potency. Through asymmetric cell division, a stem cell in adult tissues can produce an offspring that will maintain the stem cell populations and a daughter cell that will differentiate into various short-lived cell types to replace damaged or dying cells. Tumors may originate from a few transformed cancer stem cells. Stem cells have immense potential for therapeutic use in regenerative medicine and for developing anticancer therapies that specifically eliminate cancer stem cells. Germline stem cells are the source of human and animal reproduction. The knowledge gained from studying germline stem cells may find immediate application in preserving endangered wildlife, managing commercial livestock, overcoming fertility problems in humans, and treating testicular and ovary tumors. To make use of this potential, we have to learn how to isolate, characterize, and maintain germline stem cells. To isolate, characterize, and maintain stem cells, we must first understand the molecular parameters that define a germline stem cell and the mechanisms that regulate stem cell behavior. The protocols in Germline Stem Cells are intended to present selected genetic, molecular, and cellular techniques used in germline stem cell research. The book is divided into two parts. Part I covers germline stem cell identification and regulation in model organisms. Part II covers current techniques used in in vitro culture and applications of germline stem cells.

Identification and Regulation of Germline Stem Cells in Model Organisms.- The Development of Germline Stem Cells in Drosophila.- Analysis of the C. elegans Germline Stem Cell Region.- Immunohistological Techniques for Studying the Drosophila Male Germline Stem Cell.- Genetic Tools Used for Cell Lineage Tracing and Gene Manipulation in Drosophila Germline Stem Cells.- Structural Polarity and Dynamics of Male Germline Stem Cells in an Insect (Milkweed Bug Oncopeltus fasciatus).- High-Resolution Light Microscopic Characterization of Spermatogonia.- Identification and Characterization of Spermatogonial Subtypes and Their Expansion in Whole Mounts and Tissue Sections from Primate Testes.- Epigenetic Control in Male Germ Cells.- GDNF Maintains Mouse Spermatogonial Stem Cells In Vivo and In Vitro.- In Vitro Culture and Applications of Germline Stem Cells.- Ectopic Grafting of Mammalian Testis Tissue into Mouse Hosts.- Spermatogonial Stem Cell Transplantation, Testicular Function, and Restoration of Male Fertility in Mice.- Isolating Highly Pure Rat Spermatogonial Stem Cells in Culture.- Deriving Mouse Spermatogonial Stem Cell Lines.- Production of Knockdown Rats by Lentiviral Transduction of Embryos with Short Hairpin RNA Transgenes.- Testicular Germ Cell Tumors in Mice.- Study Origin of Germ Cells and Formation of New Primary Follicles in Adult Human and Rat Ovaries.

Readily reproducible protocols for the manipulation of germline stem cells in model organisms State of the art methods optimized by leading experts in germline stem cells Identification and regulation of germline stem cell in model organisms Current techniques used in in vitro culture and application in germline stem cells Presents sophisticated genetic, molecular and cellular techniques used in germline stem cell research Includes supplementary material: sn.pub/extras