The purpose of this study was to evaluate the effect of cytokines on the efficiency of gene transfer into murine hematopoietic progenitors and human K562 cells mediated by retrovirus vectors (RV) containing bacterial ...The purpose of this study was to evaluate the effect of cytokines on the efficiency of gene transfer into murine hematopoietic progenitors and human K562 cells mediated by retrovirus vectors (RV) containing bacterial neomycin-resistant (neoR) gene. The bone marrow cells were preincubated with cytokines and then transfected with supernatant containing retrovirus vectors, each for 24 h. The transfected cells were plated in the semisolid culture with or without G418. The efficiency of gene transfer into hematopoietic progenitors was estimated by biological assay and PCR analysis. The most efficient combination of the cytokines, IL-1α/IL-3/SCF,increased the efficiency of gene transfer into murine CFUGM from 6.04±1. 34% to 43. 60±5. 94%. SCF alone most efficiently facilitated the gene transfer into K562 cells from 19.04±1. 58% to 54.46±2. 13%. The results suggest that the combination of IL-1α/IL-3/SCF can increase efficiency of gene transfer into hematopoietic stem cells (HSC) and progenitors, and in the treatment of acute myeloid leukemia (AML) using autologous bone marrow transplantion (ABMT),SCF can facilitate gene transfer into hematopoietic cells in gene-marking clinical studies.展开更多
文摘The purpose of this study was to evaluate the effect of cytokines on the efficiency of gene transfer into murine hematopoietic progenitors and human K562 cells mediated by retrovirus vectors (RV) containing bacterial neomycin-resistant (neoR) gene. The bone marrow cells were preincubated with cytokines and then transfected with supernatant containing retrovirus vectors, each for 24 h. The transfected cells were plated in the semisolid culture with or without G418. The efficiency of gene transfer into hematopoietic progenitors was estimated by biological assay and PCR analysis. The most efficient combination of the cytokines, IL-1α/IL-3/SCF,increased the efficiency of gene transfer into murine CFUGM from 6.04±1. 34% to 43. 60±5. 94%. SCF alone most efficiently facilitated the gene transfer into K562 cells from 19.04±1. 58% to 54.46±2. 13%. The results suggest that the combination of IL-1α/IL-3/SCF can increase efficiency of gene transfer into hematopoietic stem cells (HSC) and progenitors, and in the treatment of acute myeloid leukemia (AML) using autologous bone marrow transplantion (ABMT),SCF can facilitate gene transfer into hematopoietic cells in gene-marking clinical studies.
