Problems in MDAMB-231 growth
Posted 15 February 2011 - 11:14 PM
I have been trying to revive MDAMB-231 using different vials frozen on different dates for over a month now without any success. The day after reviving, the cells look fine, thats when I change the media. The day after media change the cells die. I have used both DMEM with 10% serum, glutamax, antibiotics and also RPMI. The growth conditions are 37C, 5% CO2. I am sure the media is fine, because I have used them for other cell lines which are growing nicely.
I got another MDAMB-231 plate from a neighbouring lab and split and tried to grow them in my lab. But again, the cells are very unhealthy. Most of the cells are dying on the day aftr splitting and the few attached cells look unhealthy. Please Help.
Posted 16 February 2011 - 07:36 PM
Posted 16 February 2011 - 08:08 PM
Posted 17 February 2011 - 05:11 PM
What medium were the cells in when you got them? If they were in a different medium to DMEM (or RPMI), you may have to wean them onto your medium.
Posted 20 February 2011 - 11:44 PM
The confluency has been the same for over 2 weeks. Too many dead cells and the adhered cells are unhealthy. I am changing the media every alternate day.
Posted 21 February 2011 - 03:39 PM
Density problems - overconfluent when split? It doesn't sound like that should be a problem though.
Posted 23 February 2011 - 02:45 AM
And its not even getting a chance to be overconfluent. the cells are dying very fast and very few cells remain adhered to the bottom. It has been 40% confluency for over a week. Evryday I check, its the same confluency and increased no. of dead cells.
Its a complete mystery to everybody in the lab. The cells are just refusing to grow