Does anyone have experience with the N87 cell line... I am culturing it in RPMI 1640 with 20% FBS but finding the growth very slow. I was using the recommended 10% FBS but increased it, hoping to speed up proliferation. I keep getting weird jagged edged clusters that don't seem to go beyond 30-40% confluency. I'm wondering if anyone has tried a different media or added a nutrient mix to improve growth? Any tips or suggestions - much appreciated!
What dishes are you growth them on? Have you tried changing the type of plastic/ vendors? Maybe even try a fresh batch of cell
Using T75's for adherent cell lines. I've thawed multiple cryovials and they all share the same phenotype.
Hi Cassidy,
I haven't worked with that cell type but I know the problem of cells not growing properly after defreezing (I now assume that you'r etalking about the phase when you defreeze a new vial and culture it for the first time).
Do you do a cell count before transfering the cells to the flask? Is it possible that the cell density is too low (that can be a big issue for some cell lines) or the amount of viable cells isn't appropriate (maybe something happened during the stock preparation and freezing)? You could try to increase the cell density, e.g. use a smaller flask or pool two vials in one flask.
I seeded 1x10^6 cells in a T75. They do grow, just slowly. I haven't been patient enough to wait longer than 2 weeks, so the highest I've seen them is about 30% confluent. They probably would keep growing fine, I'm just impatient to run a KD experiment ;)
I would check in the literature how quick they are supposed to grow. If you should expect confluence (or at least a split-able density) with 5-7 days then it might be a problem if it takes yours 2 weeks. Some cell lines can turn strange if they're kept at too low densities or need too much time to reach confluence. I worked with Caco-2 and there a slow growth was usually a bad sign.
Maybe just make sure that of your 1x10^6 cells that were frozen not actually 50% are dead by the time you transfer them to the flask.
I've observed high concentrations of FBS the cells get stress some times is better diminish the FBS slowly since 10% to 5%.
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