The factor-dependent cell line, TF-1, established from a patient with erythroleukaemia, shows characteristics of immature erythroblasts. Addition of granulocyte-macrophage colony stimulating factor (GM-CSF) to the culture medium is required for long-term growth of the cells. Erythropoietin (Epo) can also be used to sustain TF-1 cells but for only limited periods (approximately a week). Low levels of both growth factors can act synergistically to maintain proliferation for a longer period of time than Epo alone. To eliminate the requirement of exogenous Epo for growth, TF-1 cells were co-cultured with a retroviral secreting cell line containing the human erythropoietin (hEpo) gene and a neomycin (neo) selectable marker. TF-1 cells which exhibited neo resistance (indicating infection by the retrovirus) were then grown in low concentrations of GM-CSF without the addition of Epo. Under these conditions growth of normal TF-1 cells was not sustained. The neo-resistant cells survived for more than 14 days indicating synergy between GM-CSF and the Epo synthesised by the co-cultured TF-1 cells. Radioimmunoassays performed on growth media detected concentrations up to 1 mU/ml of Epo, implying that stable integration of the retroviral vector and expression of the hEpo gene have been achieved.
|Volume||9 Suppl 1|
|Publication status||Published - Oct 1995|
- Cell Division
- Cell Line
- Cloning, Molecular
- DNA Restriction Enzymes
- DNA, Complementary
- Gene Transfer Techniques
- Genetic Markers
- Granulocyte-Macrophage Colony-Stimulating Factor
- Kanamycin Kinase
- Leukemia, Erythroblastic, Acute
- Phosphotransferases (Alcohol Group Acceptor)
- Tumor Cells, Cultured
Percy, M. J., Winter, P. C., Lappin, T. R., & McMullin, M. F. (1995). Retroviral-mediated gene transfer of the human erythropoietin gene into a factor-dependent cell line, TF-1. Leukemia, 9 Suppl 1, S66-9.