Ex vivo expansion and gene therapy using cord blood CD34+ cells.
CD34+ cells can be efficiently isolated from cord blood by sequential Percoll, Ficoll-Hypaque, and immunomagnetic bead separation. This population, containing up to 25% progenitor cells and 0.2-1% stem cells, defined by a long-term culture-initiating cell (LTC-initiating cell) assay, can be expanded in vitro in the presence of a specific combination of cytokines resulting in a > 100,000-fold increase in cells, a > 2,000-fold increase in progenitors, and an 18- to 20-fold increase in LTC-initiating cells. These latter populations can be efficiently transfected with a retroviral vector expressing a mutated dihydrofolate-reductase gene that confers methotrexate resistance.
Moore
M A
MA
SKI Cell Biology and Genetics Program, Memorial Sloan-Kettering Cancer Center, New York, NY 10021.
D015703:Antigens, CD; D018952:Antigens, CD34; D002469:Cell Separation / Q000379:methods; D002478:Cells, Cultured / Q000637:transplantation; D016207:Cytokines / Q000494:pharmacology; D005312:Fetal Blood / Q000166:cytology*; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors; D018380:Hematopoietic Stem Cell Transplantation; D006412:Hematopoietic Stem Cells / Q000166:cytology / Q000187:drug effects; D006801:Humans; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D012190:Retroviridae; D013762:Tetrahydrofolate Dehydrogenase / Q000096:biosynthesis / Q000235:genetics
D015703:Antigens, CD; D018952:Antigens, CD34; D016207:Cytokines; D011993:Recombinant Fusion Proteins; D013762:Tetrahydrofolate Dehydrogenase