Hematopoietic Stem Cell Protocols (3^rd Ed., 3rd ed. 2014) Methods in Molecular Biology Series, Vol. 1185

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

Includes cutting-edge methods and protocols

Provides step-by-step detail essential for reproducible results

Contains key notes and implementation advice from the experts

Includes supplementary material: sn.pub/extras