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Related Concept Videos

Regulation of Hematopoietic Stem Cells01:01

Regulation of Hematopoietic Stem Cells

All blood and immune cells are produced from the multipotent hematopoietic stem cells (HSCs) by the process of hematopoiesis. However, they all have a limited life span. In addition, many are depleted in immune surveillance or combatting an injury or infection. This makes blood one of the most regenerative tissues. Hematopoiesis helps replenish these blood and immune cells, restoring the body's normal functioning. However, overproduction of blood and immune cells can make them cancerous or...

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Ex Vivo Expansion and Genetic Manipulation of Mouse Hematopoietic Stem Cells in Polyvinyl Alcohol-Based Cultures
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Ex Vivo Expansion and Genetic Manipulation of Mouse Hematopoietic Stem Cells in Polyvinyl Alcohol-Based Cultures

Published on: February 10, 2023

The expanding tool kit for hematopoietic stem cell research.

William Tse1, Kevin D Bunting

  • 1Department of Medicine, Division of Hematology-Oncology, Center for Stem Cell and Regenerative Medicine, Case Western Reserve University, Cleveland, OH, USA.

Methods in Molecular Biology (Clifton, N.J.)
|March 29, 2008
PubMed
Summary

Hematopoietic stem cells (HSCs) are crucial for lifelong blood production. This overview details essential tools and methods, including transplantation assays and genetic modification, for studying HSC biology and function.

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Preparation and Gene Modification of Nonhuman Primate Hematopoietic Stem and Progenitor Cells

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Area of Science:

  • Hematology
  • Stem Cell Biology
  • Molecular Biology

Background:

  • Hematopoietic stem cells (HSCs) are vital for sustained blood cell production throughout an organism's life.
  • Significant advancements have been made in isolating pure HSCs from mouse bone marrow using cell-surface markers and functional assays.
  • In vitro methods are established for studying HSC differentiation and generating HSCs from embryonic stem cells.

Purpose of the Study:

  • To provide a comprehensive overview of the tools and methodologies available for hematopoietic stem cell research.
  • To highlight the importance of transplantation assays as the gold standard for quantitative and qualitative analysis of HSC biology.
  • To connect various research methods discussed in the second edition of Hematopoietic Stem Cell Protocols.

Main Methods:

  • Isolation of HSCs based on cell-surface phenotype and functional characteristics.
  • In vitro assays for studying hematopoietic differentiation and HSC generation from embryonic stem cells.
  • Syngeneic and xenogeneic HSC transplantation assays for quantitative assessment of repopulating function.
  • Retroviral-mediated gene transfer followed by transplantation for genetic modification of HSCs.

Main Results:

  • High purity isolation of mouse HSCs is achievable.
  • In vitro assays offer insights into HSC differentiation dynamics.
  • Transplantation assays remain the definitive method for evaluating HSC repopulating capacity.
  • Genetic modification of HSCs via retroviral vectors has significantly advanced understanding of genetic disorders, gene therapy, and leukemogenesis.

Conclusions:

  • A growing arsenal of tools exists for HSC research.
  • Transplantation assays are indispensable for quantitative and qualitative HSC analysis.
  • Retroviral gene transfer in HSCs is a powerful tool for investigating gene function and disease mechanisms.