Recently we purchased HEK-293 cell from ECACC. It is supplied at Passage 60. But HEK-293 is available at a passage say 30 elsewhere (e.g., Microbix, Canada). I am just wondering whether we have been supplied high passage cell. Has anybody used ECACC 293 cells at P-60 or above and how does it perform?
The HEK293 cell line was started in 1977 (Graham et al, J. gen. Virol. 36, 59-72 - see attachment). So I guess passage 60 is pretty good - depending at which point the count started. Most people use HEK293 cells with completely unknown passage no. Also, various sublines exist with slightly different morphologies and properties.
Low-passage cells of any non-clonal line are often more divergent, containing several distinct populations. Cell lines will usually narrow down with passage no, becoming more homogenous. Using higher passage numbers may even be an advantage, in some cases.
What is your intended application?
Thanks Alexandra. My intended application is transfection and propagation of Ad5 recombinants.
We never made Ad5, but we've tried 293T cells from various sources to make lentivirus. These lots actually differ in terms of lentiviral titer and yield (using the same protocol), although they are nominally the same cell line. Our best one we got from another lentiviral lab, with unknown passage no.
According to our experience with LV, the passage no. will not predict the performance of a specific lot of 293 cells in viral production. This is your first time with HEK293 cells, starting with cells from a certified cell bank is a good idea, to make sure they have the correct identity, and are free of contaminants. Before you start, expand the cells, and freeze a fairly large stock.
Good luck!
I am currently working with 293T cells for influenza rescue and transfection. And I found out that HEK 293 cells have various differnt passage numbers depending on the clone and subtype you are using.
I have seen a colegues who have used at lower passages and recomend that the cells to be used upto 30 passges, but this passge number is not bossible for everyone as somecells are supplied at higher passage number.
The HEK 293T I am using are from ATCC and supplied at passage 18 and I have passaged them up to p49 for 3 month in media (starting from in-house frozen vial at p23). The morphology and growh rate seems fine with consistant doubling time, eventhought in one cycle the cells were not as confluent as one would like after 3 days of incubation (4 day incubation was fine).
Nonetheless, a collegue in virology research lab who was using these cells that I have been propogating observed changes in influeza transfection at higher passages (>p30), and we are collecting data now to support this observation.
Currenty we decided to maintain the cells in media for 3 month rather than passaging to targeted passage number untill we found evidance of cell detoriaration while still rearching for further imformation about passage number for 293T/17 cells.
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