Retroviral-mediated gene transfer of the human erythropoietin gene into a factor-dependent cell line, TF-1

M J Percy, P C Winter, T R Lappin, M F McMullin

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

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.
Original languageEnglish
Pages (from-to)S66-9
JournalLeukemia
Volume9 Suppl 1
Publication statusPublished - Oct 1995

Keywords

  • Cell Division
  • Cell Line
  • Cloning, Molecular
  • DNA Restriction Enzymes
  • DNA, Complementary
  • Erythropoietin
  • Gene Transfer Techniques
  • Genetic Markers
  • Granulocyte-Macrophage Colony-Stimulating Factor
  • Humans
  • Kanamycin Kinase
  • Leukemia, Erythroblastic, Acute
  • Phosphotransferases (Alcohol Group Acceptor)
  • Radioimmunoassay
  • Tumor Cells, Cultured

Fingerprint Dive into the research topics of 'Retroviral-mediated gene transfer of the human erythropoietin gene into a factor-dependent cell line, TF-1'. Together they form a unique fingerprint.

  • Cite this