Immunomagnetic Enrichment of Human and Mouse Hematopoietic Stem Cells for Gene Therapy Applications
The hematopoietic stem cells (HSCs) constitute an ideal target for the gene therapy of inherited diseases affecting the hematopoietic system. HSCs, however, constitute a very rare population of progenitor cells, most of which are out of cycle in normal bone marrow. To facilitate their transduction with gammaretro-viral or lentiviral vectors, HSCs are generally enriched using physical or pharmacologic methods. In this chapter we describe efficient procedures which are frequently used to enrich human and mouse HSCs, aiming at the transduction of these cells with adequate gene therapy vectors or the subsequent purification of particular HSCs by fluorescence-activated cell sorting.
- Analysis of G-Proteins Regulating Signal Transduction Pathways
- Routine Culturing of ES Cells
- Reverse Transcriptase-PCR Analysis of Gene Expression in Hematopoietic Stem Cells
- Laser Ablation and Fate Mapping
- Metaphase chromosome preparation
- Practical Aspects of OCT Imaging in Tissue Engineering
- Collection of Isolated Cells for Studying Mitochondrial DNA Mutations Within Individual Cells
- Identification of Genes Involved in Tumor-Like Properties of Embryonic Stem Cells
- Production and Purification of Polyclonal Antibodies
- Detection of Histone H3 Phosphorylation in Cultured Cells and Tissue Sections by Immunostaining