Protocol Online logo
Top : Forum Archives: : Cell Biology

few cells in the cell culture - (Aug/02/2007 )


I am just beginning to culture a human uterine sarcoma cell line. This is a very fast growing cell as you now. I begun to culture about 2 weeks ago. Every time I do passage, some how there is one or two flusk with very few cells in it or cells are floating in the medium. Even after 3 days the cell cultured medium is slightly pinkish than yellow, when I compared the slow growings' with normal.

Please help me.

Thank you very much.


May be you seed too few cells in those flask.
Cell need some contact with neighbour grow properly

Hope this may help.

-Minnie Mouse-

I agree with Minnie.. I culture a cell line from Xenopus which requires cell-cell contact. If the cells are seeded too thin they almost completely stop growing and it can take over a week to reach confluency. I've also noticed that cells that were seeded thin at one point don't seem to recover and continue growing very slowly. I'd recommmend you do a few flasks with different dilutions to determine what your cells prefer and how much you need to seed to make flasks ready to split when you want.


I take it that these are HeLa cells, there shouldn't be any problems with them, they will survive a lot of mistreatment including low seeding density. I would check for bacterial contamination, including mycoplasma. If your medium (DMEM with phenol red indicator?) is changing colour quickly (e.g yellow within a day or two, or purple) or is cloudy in any way, then you will have a contamination somewhere . Get someone with cell culture experience to look at your cultures and have a look at them using a range of objectives on the microscope, are there odd looking small cells with buds coming off them?, a shimmer in the background is usually bacteria and you may be able to see chains of them under 400x magnification if you are lucky.

It could be that the batch of flasks that you are using hasn't been treated for cell culture, but then it should affect all your cultures.