Hello I'm using PC3 and DU145 cells for researching metastasis.
Before the question, please understand my poor English.
PC3 passage number is about 32 and I can't get more low passage number cell line... so I have to use this and make grows well for making cell stocks.
I usually seed PC3 on T25 flask about 60 x 10^4 for 3 days and it's about 90 x 10^4 after 3 days. I checked that doubling time of PC3 is 33 hours.
I use both RPMI and DMEM with 10% FBS and 1% P.S but in DMEM cells grow well so I'm using DMEM.
When I harvest cells, confluency was about 90% so I harvest the cells. But when I count the cells, it's about 90 x 10^4.... so it's little insufficient for my experiment. Cell morphology was good, spindle-like shape.
Can u recomment several ways to grow PC3 well? Is 15% FBS will working? Or..... do I have to buy new cells with low passage number??
Because of the insufficient cells I can't do many experiments and I'm very worried about this....
Thanks for ur help :)
Hello Hansol Cha
You should make stock of cells of earlier passage such as P3 to P10 because with higher passage number the cell phenotypic characteristics change, which may affect your experimental results.
So, I recommend that you buy cells of low passage and after culturing them for two to three passages, you may make cell stock for future use. Even though the doubling time of PC3 is 33 hours, the cell count after three days is considerably lower than expected. Using 15% FBS won't help for the growth of PC3 at P32.
If I were you, I would not prefer making cell stock at P32. In fact, for my experiment, I would thaw cells of earlier passage from the frozen stock which I would have already made.
Best.
Hi, If you want more cells. Culture in T-75, you ll get three-times the cells.
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