Abstract
We transferred new genetic information into human cell lines and hematopoietic stem cells using retrovirus vectors. These vectors, containing the dominant selectable gene (neo) for resistance to the antibiotic G418, were introduced into amphotropic helper viruses to achieve a high frequency transduction into human cells. Human cell lines or freshly obtained human marrow cells were co-cultivated with irradiated vector producing cells, and cultured in clonogenic assays with and without G418. Survival curves of non-transfected cells with increasing concentrations of G418 resulted in a dose-dependent inhibition of colony growth with a final dose of 500 to 800 μg/ml. Transfected promyelocytic leukemia (HL-60), erythroleukemia (OCI-M2) and different lymphoid cell lines with N2 and SV(X) vectors, revealed G418 resistant colonies in frequencies ranging from 3.2 to 44%. Similar frequencies were found with human marrow cells. Transfection with either N2 or NEOμ vectors produced G418 resistant hematopoietic progenitor colonies of the erythroid (BFU-E) granulocyte-macrophage (CFU-GM), and mixed (GFU-GEMM) lineages in frequencies ranging from 8 to 40% after 14 days in culture. We conclude that selected retrovirus vectors can be successfully used to insert new genetic information into pluripotent and committed human hematopoietic progenitor cells.
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