Help with BT549 Morphology, Maintenance & Possible Contamination - (Jan/07/2012 )

Hi,

I'm currently an undergrad student working on BT549 & MB231 to study signalling pathways. However, I've encountered much difficulty in maintaining sufficient and healthy BT549 for my work.

Growth Characteristic
Does anyone have any idea what the doubling time of BT549 is? My senior told me it was similar to MB231, which is 36H. But ATCC does not mention this, so I was hoping if someone could verify this for me. One of the growth characteristics I noticed with my BT549 is that they require a very high seeding density in order for them to grow well (confluent T75 = 2xT75 or less), have anyone experienced this problem as well? The BT549 I'm growing also seem to slow down a lot in their growth towards the later passages, and I'm not sure if this is due to genetic instability of the cell line, maintenance issues or contamination, could someone help verify on this?

Morphology
I was doing a search online to find images of what BT549 should look like for my project that requires me to work with this cell line. Apparently ATCC does not have an image to show what it's morphology is like, but only describes that it should look like epithelial cells.

Maintenance
Although ATCC recommends the supplementation of insulin for the maintenance of BT549, the lab I'm working in does not use insulin to maintain this cell line. The growth characteristic of the BT549 that we maintain seem to look like elongated versions of MB-231, with a very thin and needle-like body, not sure if it is suppose to be this way. I'll try to get a picture my the BT549 and upload it soon.

Also, in the course of maintaining BT549, i realized that a lot of black dots seem to appear in the back ground. Not sure if this is a characteristic of this cell line or contamination. But it could well be cell debris moving in the background through Brownian motion. I'll try to get an image of this up soon as well.

Much help is appreciated. Thanks.

Regards,
Unknownko

BT549 doubling time is in 1-2 days usually, that is definately more rapid than MB-231 , I work with both cell lines. These cells grow very rapid and very well in right culture conditions. Are you using RPMI with glutamax from invitrogen, sub with 10% FBS and 0.5% pen strep. You do not need insulin. These cells look almost identical to MB-231.