Hematopoietic Stem Cell Protocols

The hematopoietic stem cell (HSC) field has rapidly grown in the past several years as new technologies have been developed and the older tried and true methods have been used in new ways. Major advances in the isolation of HSC and progenitor subsets have been sparked by the gene expression array technologies, leading to the identification of new markers for prospective isolation. The first edition of Hematopoietic Stem Cell Protocols is a very thorough resource covering historically the basic techniques that have been published extensively over many years. To put together, a follow-up to this excellent book has been a hard act to follow. The intent of this book is to be distinct and not duplicate but to build on the strong basic assays and include updated approaches for the combination of phenotypic and functional analyses. This second edition of Hematopoietic Stem Cell Protocols thus aims to provide timely protocols developed in the HSC field since the last comprehensive review in this series. The approaches described in this book should be readily useful for the basic science researcher, especially those familiar with the use of mouse genetic models. The chapters in this book are geared toward the laboratory-based researcher and the development of pre-clinical studies to apply HSCs toward treating blood diseases more effectively.

Part I. Overview 1. The Expanding Tool Kit for Hematopoietic Stem Cell Research William Tse and Kevin D. Bunting Part II. Stem Cell Enrichment and Analysis 2. Isolation of Quiescent Murine Hematopoietic Stem cells by Homing Properties Tarja A. Juopperi and Saul J. Sharkis 3. Mobilization of Hematopoietic Stem and Progenitor cells (HSPC) in Mice Simon N. Robinson and Ronald P. van Os 4. Transmigration of Human CD34+ cells Seiji Fukuda and Louis M. Pelus 5. Flow Cytometry Based Cell Cycle Measurement of Mouse Hematopoietic Stem and Progenitor Cells Hongmei Shen, Matthew Boyer, and Tao Cheng 6. Analysis of Apoptosis in HSC by Flow Cytometry William G. Kerr Part III. In vitro Assays and Differentiation 7. In vitro Hematopoietic Differentiation of Murine Embryonic Stem (ES) Cells Jinhua Shen and Cheng-Kui Qu 8. Hematopoietic Development of Human Embryonic Stem Cells in Culture Xinghui Tian and Dan S. Kaufman 9. In vitro Human T Cell Development Directed by Notch-ligand Interactions Génève Awong, Ross N. La Motte-Mohs and Juan Carlos Zúñiga-Pflücker 10. In vitro Assays for Cobblestone Area Forming Cells, LTC-IC, and CFU-C Ronald van Os, Bertien Dethmers-Ausema, and Gerald de Haan Part IV. Transplantation Assays for Mouse and Human HSC 11. HSC Transplant in Mice by Intra Femoral Injection Yi Zhao and Yuxia Zhan 12. Hematopoietic Stem Cell Transplant into Non-myeloablated W/Wv mice to Detect Steady-state Engraftment Defects Zhengqi Wang and Kevin D. Bunting 13. Isolation and FunctionalCharacterization of Side Population Stem Cells Jonathan B. Johnnidis and Fernando D. Camargo 14. Transplantation of Chimeric Fetal Liver to Study Hematopoiesis Sigrid Eckardt and K. John McLaughlin 15. Immunodeficient Mouse Models to Study Human Stem Cell-mediated Tissue Repair Ping Zhou, Sarah Hohm, Ben Cappocia, Louisa Wirthlin, David Hess, Dan Link, and Jan Nolta Part V. Genetic Modification of HSCs and Imaging Engraftment 16. Retroviral Transduction of Murine Hematopoietic Stem Cells Peter Haviernik, Yi Zhang, and Kevin D. Bunting 17. Lentiviral Gene Transduction of Mouse and Human Stem Cells Zhaohui Ye, Xiaobing Yu, and Linzhao Cheng 18. Retroviral Integration Site Analysis in Hematopoietic Stem Cells Olga S. Kustikova, Christopher Baum, and Boris Fehse 19. A Detailed Protocol for BAC Recombineering to Study Essential Genes in Stem Cells Andriy Tsyrulnyk and Richard Moriggl 20. Bioluminescence Imaging of Hematopoietic Stem Cell Repopulation in Murine Models Yuan Lin, Joe Molter, Zhenghong Lee, Stanton L. Gerson