Different cell culture plastics, problems with cell adherance - (Oct/02/2008 )
We are having some problems with our cancer cells properly adhering to our cell culture flasks. We are using Sarstedt flasks and plates, and cell adherence is not consistent. For example in my latest experiment to do some time-course treatment of my cancer cells, 8 of 12 flasks properly adhered and 4 of the flasks contained cells that were rounded or significantly lower number of cells. I make a master mix of cells so all the flasks got the same media, and the same cells all from the same master mix. So the variation has to be coming from the flasks themselves. Has anyone had similar problems such as this? I've been having to make some extra flasks of cells as some of the flasks always seem to not work out too well.
I'm also considering switching to a different brand of flasks, and I've heard good things about Corning and BD falcon and some of my colleagues that use Corning say that they never have a problem.
Even primary cells or cells that have a high adherence factor have inconsistent binding to the flasks, despite being split from a master mix of trypsinized cells.
I've also heard that rinsing the flasks with PBS prior to adding cells to the flasks can reduce cell binding variability. Is there any truth to that??
-Rubyeye
I'm also considering switching to a different brand of flasks, and I've heard good things about Corning and BD falcon and some of my colleagues that use Corning say that they never have a problem.
Even primary cells or cells that have a high adherence factor have inconsistent binding to the flasks, despite being split from a master mix of trypsinized cells.
I've also heard that rinsing the flasks with PBS prior to adding cells to the flasks can reduce cell binding variability. Is there any truth to that??
-Rubyeye
Are the flasks specifically treated for tissue culture? Also, if you are trypsinizing the cells, are you inactivating the trypsin after they have been treated? Although I don't normally rinse my flasks or plates with PBS, I wouldn't think that would have any effect an the ability of the cells to adhere. I use Corning plates and flasks, and have never had a problem with them.
I think Corning flasks and plates are better quality than Sarstedt ones.
How good is your incubator? Is there a pattern as to which flasks failed to attach based on where they were placed in the incubator?
We were just able to test some corning flasks and our technician reported that the cells adhered much better than with sarstedt flasks. Also the level of confluency was different and was the level was more consistant within corning flasks. We also have Some BD Falcon flasks to test and will see if they are better than sarstedt.
Hello,
I also had some of problems with cell adherence. I've tried both BD Falcon brand culture flasks and Corning CellBIND Surface culture flasks. The Falcon flasks were okay, but the Corning CellBIND flasks worked alot better.
Also, try coating the plastic with collagen (or some other ECM component). It has helped my cells to attach amazingly well!
You can try requesting free samples from Corning on their website:
http://www.corning.com/lifesciences/whats_...requestform.asp
Hope this info helps!
I've attached an image of the result of the test.
BD falcon flasks were uniform in the confluency of the cells and the sarstedt flasks were not uniform. 3 of 6 of the sarstdet flasks had low attachment while the other 3 had some moderate attachment. All of the BD falcon flasks had better attachment. These flasks were plated from a master mix of cells and were cultured for 2 days at 37 degrees C, 95% humidity and 5% CO2. The cell type is 4T1 mammary cancer cells.
Update: Corning has agreed to send us some samples so we'll be testing those as well. Hopefully we can decide on a brand soon!