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


Autoria(s): Percy, M J; Winter, P C; Lappin, T R; McMullin, M F
Data(s)

01/10/1995

Resumo

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.

Identificador

http://pure.qub.ac.uk/portal/en/publications/retroviralmediated-gene-transfer-of-the-human-erythropoietin-gene-into-a-factordependent-cell-line-tf1(4c0d7dc9-c138-44f5-9294-f4174439734a).html

Idioma(s)

eng

Direitos

info:eu-repo/semantics/restrictedAccess

Fonte

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 , vol 9 Suppl 1 , pp. S66-9 .

Palavras-Chave #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
Tipo

article