I have tried many times to setup cultures using this cells line (SW840) in 10 ml DMEM + 5 ml FBS + 500μl P/S and then incubate the culture in Co2 incubator at 37 c and 5 % of co2, but the density stills low!!! Any suggestion?
Hi Fadwa .
Are you saying you set up cells in 10 ml DMEM containing 5 ml of FBS?
That is a very high amount of media: FBS is generally used at ~ 10% (9 ml DMEM + 1 ml FBS). Depending upon the concentration P?S is generally used at i.e. 1:100 so this might also be too high.
I personally have not used these cells (I have grown other Duke cells though).
Where did you get these cells from?
ATCC recommends that they are grown in L15 media (which should not be placed in a 5% CO2 environment).
http://www.atcc.org/products/all/CCL-228.aspx#culturemethod
Another reason cells fail to grow is due to the quality of the FBS used. Some cell lines are quite sensitive to this (and this is why FBS is often batch tested against a particular cell line).
Sorry I can't be more help,
Gary
IF it is 5ml FBS in 10 ml media, then yes its too high as Gary said. The recommended concentration is 10 percent FBS. Also the recommended concentration of penstrep is 0.5-1 ml in 100 ml media; this too is high in your case. I have cultured SW480 cells in DMEM using 10 percent FBS and 5ml penstrep in 500ml media. It has fairly high growth rate.
Thank you all . I think I should re do it again following your recommendations
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