Culture medium for cell growth and transfection
View Patent ↗The present invention is directed generally to cell culture media useful for introducing macromolecules and compounds (e.g., nucleic acid molecules) into cells (e.g., eukaryotic cells) in the presence of said media. Cells containing introduced materials can be further cultured in the media. In particular, the invention allows introduction of nucleic acid molecules (e.g., vectors) into cells (particularly eukaryotic cells) and expression of proteins encoded by the nucleic acid molecules in the cells. The invention obviates the need to change the cell culture medium each time a different procedure is performed with the cells (e.g., culturing cells vs. transfecting cells). The invention thus provides efficient and high throughput methods to transform/transfect culture and cells avoiding the need for multiple manipulations and transfers of cells during transfection and expression studies.
1. A method for introducing a nucleic acid into a 293F cell under serum-free suspension culture conditions, said method comprising:
(a) culturing the 293F cell in the serum-free conditions comprising serum-free cell culture medium, wherein the cell culture medium supports the introduction of the nucleic acid into the 293F cell and also supports cultivation of the 293F cell subsequent to introduction of the nucleic acid, wherein the nucleic acid encodes a protein and is operably linked to a promoter;
(b) introducing a lipid aggregate comprising the nucleic acid into the cell culture medium in which the 293F cell is cultured under conditions sufficient to introduce the nucleic acid into the 293F cell; and
(c) cultivating the 293F cell into which the nucleic acid has been introduced in the culture medium under conditions sufficient to express the protein from the nucleic acid,
wherein during step (b) the culture medium is not replenished, replaced or supplemented for at least 48 hours; and
wherein cultivation of the 293F cell in step (c) is accomplished in a volume of the culture medium that is about the same or up to about 2 times the volume of the culture medium in which step (b) occurred.
2. The method of claim 1 , wherein the lipid aggregate comprises a cationic lipid.