SpringerProtocols: Abstract: 3. Murine Hematopoietic Stem Cell Transduction Using Retroviral Vectors

3. Murine Hematopoietic Stem Cell Transduction Using Retroviral Vectors

By: Ute Modlich³, Axel Schambach³, Zhixiong Li³, Bernhard Schiedlmeier³

Abstract

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Hematopoietic stem cells (HSCs) represent an important target cell population in bone marrow transplantation and gene therapy applications. Their progeny cells carry the genetic information of the HSCs and replenish the blood and immune system. Therefore, in the setting of inherited diseases, transduction of HSCs with retroviral vectors (including gammaretro- and lentiviral vectors) offers the possibility to correct the phenotype in all blood lineages as demonstrated in clinical trials for immunodeficiencies (e.g., X-SCID). In the process of developing gene therapy strategies for patient applications, suitable mouse models for the human gene therapy are important to validate the concept. Stem-cell-enriched populations such as lineage negative cells as the functional equivalent of human CD34⁺ cells can be isolated from murine bone marrow and efficiently transduced using retroviral vectors. This chapter provides a step-by-step protocol for retroviral transduction of murine lineage negative cells.

Affiliation(s): (3) Department of Experimental Hematology, Hannover Medical School, Hannover, Germany

Book Title: Genetic Modification of Hematopoietic Stem Cells: Methods and Protocols

Series: Methods in Molecular Biology | Volume: 506 | Pub. Date: Jul-01-2008 | Page Range: 23-31 | DOI: 10.1007/978-1-59745-409-4_3

Subject: Cell Biology

Key Words: Hematopoietic stem cells - Retroviral vector - Lentiviral vector - Gene therapy - Transduction - Cell culture - Serum-free medium - Cytokines

References

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