Sk-Br-3 cells - (Sep/19/2006 )

Does anybody have any experience with growing Sk-Br-3 cells? We are having serious trouble getting them to grow under the conditions specified by ATCC. we have tried numerous times but no luck. Any ideas??

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What is the medium you use?

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We are currently using McCoy's 5A with 15% FBS and as of today we have a layer of something that looks more like debris than cells. We have also tried RPMI 1640 10% FBS and alpha MEM 10% FBS. We have used cells from different sources but none of them appear to be happy (all our other cell lines are growing beautifully........we do know how to do this stuff usually).

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I can't tell what your problem is: could be the cells that you thawed. Is the vial from ATCC?
Here's my experience: I thawed a vial on Thursday and put in T75 flask and it was ~80% confluent on Friday. So I had to expand it to T175 X 2 on Friday - that is day 2 itself.
I used: DMEM with high Glucose + 10% FBS + 1% L Glutamine + 1% pen/strep.

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Mccoy 5A with 10% FBS and glutamine (7.5 mM)

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We culture our SK-Br-3 using RPMI supplemented with 10% FBS, 2% pen/strep, 1% sodium pyruvate, and 1.25% L-glut.

We usually seed a T-75 with 5x10^4 to 7.5x10^4 cells/ml and culture them 2x a week... usually after 72 and 96 hours.

use 5ml trypsin/EDTA (for a T-75) for no longer than 9 mins than add 20 ml supplemented RPMI.

Ive been culturing these cells for the past year with no problems. hope this helps

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