Klug / Jordan Hematopoietic Stem Cell Protocols

Erscheinungsjahr 2008
ISBN: 978-1-59259-140-4
Verlag: Humana Press
Format: PDF
Kopierschutz: 1 - PDF Watermark

E-Book, Englisch, 332 Seiten

Reihe: Springer Protocols

The ability to highly purify and characterize hematopoietic stem cells (HSC) from mice and humans has opened up an exceedingly rich field of basic science research with enormous clinical potential. Many of the techniques used in st- ies of HSC biology have become more standardized over the last several years, which makes it possible to compile a set of methods that can be used by both seasoned investigators and novices in the stem cell field. We have attempted to be as comprehensive as possible and yet focus on what we perceive to be the most widely used approaches for studies of murine and human HSC. This first edition of Hematopoietic Stem Cell Protocols will therefore have some obvious omissions that were dictated by contemporary circumstances. It is our hope that readers will feel free to contribute their personal suggestions for further chapters as well as on how existing chapters can be improved for future editions. We certainly expect that old approaches will be refined, new assays will be developed, and other animal model and vector systems will be described that will become the new gold standards for future work. Our s- cere thanks goes out to all of the contributors and to those in the stem cell field that have enlarged our thinking and provided new tools to further understand this fascinating cell type.

Zielgruppe

Professional/practitioner

Autoren/Hrsg.

Klug, Christopher A.

Jordan, Craig T.

Inhaltsverzeichnis

Isolation and Analysis of Hematopoietic Stem Cells from Mouse Embryos.- The Purification of Mouse Hematopoietic Stem Cells at Sequential Stages of Maturation.- Flow Cytometry and Immunoselection of Human Stem Cells.- Method for Purification of Human Hematopoietic Stem Cells by Flow Cytometry.- Noninvasive Measurement of Hematopoietic Stem Cell Cycling and Turnover by In Vivo Bromodeoxyuridine Incorporation.- Isolation and Characterization of Primitive Hematopoietic Cells Based on Their Position in the Cell Cycle.- Hematopoietic Colony-Forming Cells.- Long-Term Culture-Initiating Cell Assays for Human and Murine Cells.- The Cobblestone-Area-Forming Cell Assay.- CFU-S.- Intrathymic Injection for Analysis of T-Cell Progenitor Activity.- Quantitation of Murine and Human Hematopoietic Stem Cells by Limiting-Dilution Analysis in Competitively Repopulated Hosts.- Ex Vivo Expansion of Human and Murine Hematopoietic Stem Cells.- Hematopoietic Development of ES Cells in Culture.- Genetic Modification of Murine Hematopoietic Stem Cells by Retroviruses.- Retroviral Transduction of FACS-Purified Hematopoietic Stem Cells.- Retroviral-Mediated Transduction and Clonal Integration Analysis of Human Hematopoietic Stem and Progenitor Cells.- Production of Lentiviral Vector Supernatants and Transduction of Cellular Targets.- Reverse Transcriptase-PCR Analysis of Gene Expression in Hematopoietic Stem Cells.- Identification of Differentially Expressed Genes in Sorted Cell Populations by Two-Dimensional Gene-Expression Fingerprinting.