Cells growing very slowly - (May/14/2012 )
I am working with Fetal Bovine Aortic endothelial cells, from the information on the internet it looks like its doubling time is 16 hrs. But even after keeping it for more than 48 hours I don't see much growth.
Since these cells weren't growing real fast, I seeded them at a very high density. Could you please suggest changes in conditions or possible reasons for the slow cell growth.
I am using DMEG medium with 10% FBS and the medium appears to be fine because I cultured mink lung cells and they are happily growing.
I would appreciate any suggestions.
decelerated growing of cells is difficult to amend if you have no idea because it could be anything: quality of cells, wrong cells, too dense seeding, wrong plates, bad medium, bad atmosphere/humidity/temperature conditions, infections etc
Have you checked your cells for mycoplasma ?
Thanks for your response.
As I had mentioned that I am using the same medium to culture another cell line and they are seem to be dividing normally so I think it shouldn't be a problem with media or cell growing conditions.
I haven't checked for mycoplasma contamination but thats a good idea, I shall check and see.
Just saying, but 16 h is incredibly fast for a primary cell type, even HeLa only double about every 18 h. I wouldn't be surprised if you have your numbers wrong there; the papers I found indicate that 35-82 hours doubling time, depending on passage.
Those media which have https://en.wikipedia.org/wiki/Sodium_pyruvate are better.
And for removing Mycoplasma infection
In fact, normocin is used to prevent mycoplasma proliferation, but not to treat it. There are other antibiotics, which "supposedly" get rid of mycoplasma, but frankly, I think they will also select for a certain subpopulation of your cells, since they appear to be quite toxic. I would recommend throwing out the cells, medium and everything, sterilizing incubator and hood, and opening a new batch of cells.
Problem is that it may not be a mycoplasma contamination -- virus or intracellular bacteria, which is quite tough to deal with.