Stable Cell Line Transient Transfection

When developing a stable transfection researchers use selectable markers to distinguish transient from stable transfections.

Stable cell line transient transfection.

In a subpopulation of transfected cells whether the desired effect is a stable or transient transfection the transfected genetic material will integrate into the genome. Stable transfection is the long term introduction of foreign dna into cells. In transient transfection the transfected material enters the cell but does not get integrated into the cellular genome. In order to create stable cell lines investigators will take advantage of this natural occurrence and introduce the gene of interest along with a selectable marker.

Stably transfected cells pass the foreign dna through progeny. Transient transfection of mammalian cells has been employed for recombinant protein expression since the invention of transfection reagents the ability to express milligram to gram amounts of recombinant protein has relied mainly on the creation of stable cell lines. Typical cell density ranges are as follows. Unlike transient transfection in which introduced dna persists in cells for several days stable transfection introduces dna into cells long term.

However stable inheritance can be observed in nongenomic dna as well. For each individual stable cell line to be created plate cells in three t75 flasks and one 6 well tissue culture plate approximately 18 24 hours before transfection such that they reach high confluence 60 80 at the time of transfection. The plasmid dna or other type of nucleic acid typically has a reporter gene that allows a scientist to monitor the expression of the reporter gene usually within 1 2 days post transfection. 0 8 2 4 10 5 cells ml of complete media.

Transfection can be classified into main two types named transient transfection and stable transfection during transient transfection the gene of interest fails to integrate with the host genome and is expressed temporally. Key difference transient vs stable transfection transfection is a process which is involved in the gene transfer of eukaryotic cells using chemical or physical methods. Descendants of these transfected cells therefore will also express the new gene resulting in a stably transfected cell line. Unlike the short term protein expression observed using transient transfection approaches generating cell lines using lentiviral vectors enables long term protein expression studies.

Stably transfected cells pass the introduced dna to their progeny typically because the transfected dna has been incorporated into the genome but sometimes via stable inheritance of nongenomic dna. Therefore to make stably transfected cell lines foreign dna has to be integrated into the genome of the cell line. One is achieved by eukaryotic vectors that harbor elements for episomal maintenance in the nucleus of a transfected cell.