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Updating and building upon previous editions, Hematopoietic Stem Cell Protocols, Third Edition provides up-to-date protocols from leading stem cell researchers. This in-depth volume presents a clear view of the landscape of assays available to the stem cell researcher working in the growing hematopoietic stem cell (HSC) field. A robust and active field, it is supported by an abundance of innovative mouse models and molecular tools for analysis of phenotypes and functions in mouse and human cells. Understanding more about hematopoietic stem cell biology is integral if these versatile cells are to be applied effectively to treat and cure a wide range of blood diseases. An introductory chapter puts the major contributions of the book into the proper perspective. Written in the 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 protocols, and notes on troubleshooting and avoiding known pitfalls.
Essential for the laboratory-based researcher, Hematopoietic Stem Cell Protocols, Third Edition is a much needed technical resource in the critically important field of hematopoietic stem cell investigation.
Part I. Overview
1. The Hematopoietic Stem Cell Landscape
Kevin D. Bunting and Cheng-Kui Qu
Part II. Stem Cell Enrichment and Heterogeneity
2. Investigating the Interaction Between Hematopoietic Stem Cells and Their Niche During Embryonic Development: Optimizing the Isolation of Fetal and Newborn Stem Cells From Liver, Spleen, and Bone Marrow
Huimin Cao, Brenda Williams, and Susan K. Nilsson
3. Single-Cell PCR Profiling of Gene Expression in Hematopoiesis
Jose Teles, Tariq Enver, and Cristina Pina
4. Hematopoietic Stem and Progenitor Cell Mobilization in Mice
Jonathan Hoggatt, Tiffany A. Tate, and Louis M. Pelus
5. Cell Cycle Measurement of Mouse Hematopoietic Stem/Progenitor Cells
Brahmananda Reddy Chitteti and Edward F. Srour
6. Flow Cytometry Analysis of Signaling and Apoptosis in Hematopoietic Stem Cells
Lei Dong and Cheng-Kui Qu
Part III. Molecular and Cellular Characterization
7. Gene Expression Profiling of Hematopoietic Stem Cells (HSC)
Nalini Raghavachari
8. Measuring MicroRNA Expression in Mouse Hematopoietic Stem Cells
Wenhuo Hu and Christopher Y. Park
9. DNA Methylation Profiling of Hematopoietic Stem Cells
Amber H . Begtrup
10. Metabolic Characterization of Hematopoietic Stem Cells
Fatih Kocabas, Junke Zheng, Chengcheng Zhang, and Hesham A. Sadek
11. Nano Proteomic Assays on Hematopoietic Stem Cells
Heath L. Bradley, Himalee Sabnis, Deborah Pritchett, and Kevin D. Bunting
Part IV. In vitro Assays and Differentiation
12. Hematopoietic Differentiation of Pluripotent Stem Cells in Culture
Jason A. Mills, Prasuna Paluru, Mitchell J. Weiss, Paul Gadue, and Deborah L. French
13. Ex vivo Assays to Study Self-Renewal, Long-Term Expansion and Leukemic Transformation of Genetically Modified Human Hematopoietic and Patient-Derived Leukemic Stem Cells
Pallavi Sontakke, Marco Carretta, Marta Capala, Hein Schepers, and Jan Jacob Schuringa
14. Ex vivo Expansion of Murine and Human Hematopoietic Stem Cells
Phuong L. Doan and John P. Chute
15. Using Microfluidics to Investigate Hematopoietic Stem Cell and Microniche Interactions at the Single Cell Level
Byungwook Aha, Zhengqi Wang, David R. Archer, and Wilbur A. Lam
Part V. Transplantation Assays and Imaging Engraftment
16. Five Lineage Clonal Analysis of Hematopoietic Stem/Progenitor Cells
Ryo Yamamoto, Yohei Morita, and Hiromitsu Nakauchi
17. Intravital Imaging of Hematopoietic Stem Cells in the Mouse Skull
Juwell W. Wu, Judith M. Runnels, and Charles P. Lin
18. Immune Deficient Mouse Model for Human Hematopoietic Stem Cell Engraftment and Immune System Development
Ken-Edwin Aryee, Leonard D. Shultz, and Michael A. Brehm
19. Homing and Migration Assays of Hematopoietic Stem/Progenitor Cells
Xi C. He, Zhenrui Li, Rio Sugimura, Jason Ross, Meng Zhao, and Linheng Li
Part VI. Genetic Modification
20. Retroviral Transduction of Murine and Human Hematopoietic Progenitors and Stem Cells
Marioara F. Ciuculescu, Christian Brendel, Chad E. Harris, and David A. Williams
21. Lentiviral Gene Transduction of Mouse and Human Hematopoietic Stem Cells
Niek P. van Til and Gerard Wagemaker
22. High Throughput Genomic Mapping of Vector Integration Sites in Gene Therapy Studies
Brian C. Beard, Jennifer E. Adair, Grant D. Trobridge, and Hans-Peter Kiem
23. Barcoded Vector Libraries and Retro- or Lentiviral Barcoding of Hematopoietic Stem Cells
Leonid V. Bystrykh, Gerald de Haan, and Evgenia Verovskaya
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