DNA transfection in COS cells: a low-cost serum-free method compared to lipofection.
We describe a defined medium that allows efficient DNA transfections in COS cells and transient expression of the corresponding recombinant protein in serum-free conditions. With a modified DEAE-dextran/chloroquine method, we obtained 80% more transfected cells expressing the recombinant human interleukin-2 receptor than with transfection with cationic liposomes, one of the most efficient techniques to date. The absence of serum in the culture medium should reduce subsequent purification steps for production of recombinant mammalian proteins. Moreover, it should allow investigations dealing with the role of serum or other exogenous factors on mRNA stability or post-translation events during protein synthesis.
Laboratoire C.N.R.S., I.C.I.G. Hôpital Paul-Brousse, Villejuif, France.
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JP
Lévesque
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Sansilvestri
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Hatzfeld
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Hatzfeld
D002460:Cell Line; D002738:Chloroquine; D003365:Costs and Cost Analysis; D016895:Culture Media, Serum-Free; D003637:DEAE-Dextran; D004247:DNA / Q000235:genetics; D006801:Humans; D008081:Liposomes; D010957:Plasmids; D015375:Receptors, Interleukin-2 / Q000235:genetics; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics; D014162:Transfection
D016895:Culture Media, Serum-Free; D008081:Liposomes; D015375:Receptors, Interleukin-2; D011994:Recombinant Proteins; D002738:Chloroquine; D004247:DNA; D003637:DEAE-Dextran