The transfection method determines the cell type for stable integration.
Stable cell line generation ppt.
Introduction based on many years experience on cell line development creative biogene s cell culture scientists focus on optimizing all parameters specific to your cell lines to efficiently produce stable cell lines according to your requirements.
Protein overexpressing cell lines rnai knockdown cell lines and reporter stable cell lines luciferase gfp rfp yfp.
Initially we use 400 ug ml g418 in the media when selecting for stable clones.
Download altogen labs stable cell line development powerpoint presentation.
Stable cell lines expressing a protein of interest or shrna constructs targeting gene of interest are important research tools in life science widely used in drug discovery and compound screening programs.
24 to combine the advantages conferred by employing a stable vector producing cell line with the enhanced biosafety properties of ppt deleted vectors we chose to stably transfect a ppt deleted vector.
Major challenges for generation of stable cell lines are low transfection efficiency and or integration frequency.
So far generation of stable cell lines has been a major challenge for many cell types e g jurkat mcf7 or u937 since overall transfection efficiencies and or integration frequencies have been low.
The amount of g418 required to kill cells not expressing the construct will vary from cell line to cell line.
Stable expression can be influenced by the transfection method used.
Methods to generate stable cell lines 01 02 a mixed population of drug resistant cells generate a monoclonal cell line a mixed population of drug resistant cells can be used directly for experimental analysis with the advantage of generating fast results but also the disadvantage of dealing with an undefined and genetically mixed cell population.
Further generating stable cell lines or performing large scale or multiple small scale transient transfections to serve as a continuous source of recombinant mab for screening and preclinical development is impractical for dozens or hundreds of lead mabs.
Consequently both integrase mediated integration of ppt deleted iclvs and illegitimate integration of ppt deleted idlvs are severely impaired.
While common transfection methods such as lipofection can be used for the stable expression in easy to transfect cell lines e g hela cos 7.
Altogen labs offers several stable cell line generation services.
An ideal mab cloning and expression platform would contain a single high throughput.
We have titrated the amount required for our hela cell lines and this can be a starting point for other cell lines.
A systematic and comprehensive guideline for generation of stable cell lines including the background and operations at each step.
