For the cell culture, I am using tissue culture plates; I just noticed that the ones we received recently have this label'' Non tissue culture treated plate''. Is there sth related to evaporation? what else? Any way, I found out that cells in the well plates even in 24 wells with a very low concentration like 500 cells/ml get really clumpy while they are not in this way when they grow in a bigger space like flasks. Do you think it's better to use flasks instead of well plates since I need to work on cell structures?
Best,
"Cell culture-treated" usually refers to the process by which otherwise hydrophobic polystyrene surfaces are made to become hydrophilic, usually by increasing negative charge through a chemical means. This increased negative charge is important for cell attachment, and may explain why cells cultured in your non-treated plasticware clump (namely that they are not attaching adequately to the surface). What cell type are you using?
"Cell culture-treated" usually refers to the process by which otherwise hydrophobic polystyrene surfaces are made to become hydrophilic, usually by increasing negative charge through a chemical means. This increased negative charge is important for cell attachment, and may explain why cells cultured in your non-treated plasticware clump (namely that they are not attaching adequately to the surface). What cell type are you using?
1. "Non-tissue culture treated" surface is more hydrophobic than "tissue-culture treated" surface and is similar to the surface of a bacteriological Petri dish.
2. "Tissue-culture treated" is for cell culture of anchorage-dependent cells and "non-treated" is for suspension cell culture applications, such as mammalian cells that grow in suspension and bacterial cell culture where attachment is not required.
I would agree with Joshua. For any kind of cultured cells, the cells will not attach properly if you do not use the "cell culture treated" or the chemically treated plastic ware. So, while changing media, you could lose a lot of cells unless you use a centrifuge to precipitate the cells. Adherence to the plastic surface is also important for the growth of the cells (adherent types). For example, the cell culture treated dish might give you twice the number of cells than cells from a non-treated dish. We know this as a tech in the lab made a mistake of buying the non-treated dishes once!
Thank you so much for your answers. The cells I'm working with are BY2 cells( a very fast growing plant cell line) and I'm using the suspension culture. Because I need many replicates for different treatments, I tried tissue culture wells. However, I got to know that cells get clumpy in non treated tissue culture while I didn't try treated tissue culture well plates. If treated tissue culture wells are just for adhering, I don't need it. But probably they can help the shaking to be better. Am I right?
Thank you,
You're right, if your cell line grows in suspension culture, it may even be preferable to have them in non-treated wells so that cells do not 'stick' to the bottom (as Yuan-Yeu mentioned above). In this event, your observation that the cells clump in a dish but not in the flask (at identical concentration and medium depth, i presume) is even more mysterious. Are there any other differences in the handling of the cell culture vessels that you can think of? I know a common mistake for people who aren't used to using circular culture vessels is swirling the dish to mix - this actually results in cells being pulled to the center of the dish and clumping together, particularly a problem for adherent cells. Think for awhile about potential variations in your procedure and you may find something you do differently that explains the clumping. Good luck!
Dear Joshua, Thanks. Actually, I' thinking the problem might be mostly because of shaking since to decrease the evaporation, I use a box with some water underneath of the plates inside the box. So, the plates are not in a good position for shaking probably while the flask are directly on the surface of the shaker and, shake well.
Regards,
Plasticware that is NOT meant to be used for tissue culture has sometimes been sterilized with ethylene oxide. Outgassing of residual ethylene oxide can be toxic for cells.
So even for a suspension cell line, it's better to use TC treated plates/dishes?
Due to the possible toxicity of the residues after the sterilization process?
Thanks
No. There is no advantage to using TC plastic for non-adherent cell lines as long as one stays clear of ethylene oxide sterilized plasticware. Look for plasticware that has been sterilized by gamma radiation. Even better, look for plasticware from Greiner that is produced by a clean manufacturing process and does not require sterilization. It works very well for non-adherent cells.
Mahbobeh zamani babgohari is dealing with BY-2 cell lines, which is a plant cell line (tobacco). And he is doing plant cell suspension culture. So, 'tissue-culture-treated' plates are not needed. But, I am wondering that whether the BY-2 cells will stick to the well walls of the 'tissue-culture-treated' plates if he uses them? Plant cells have cell walls, not like mammalian cells which have no cell walls.
Hi Mahbobeh, I think the problem is that sometimes cells would want to attach to something to grow, and TC treated plates provide better attachment. Here's a link from sigma with an interesting story on cell plate surfaces:
http://www.sigmaaldrich.com/technical-documents/articles/biofiles/evolution-of-cell.html
In their results, the super-hydrophilic (specially TC treated) plates yields a better cell growth than regular TC plates. Perhaps your cells would want some attachment also?
Tissue culture plates are treated so that the cells will adhere and have nothing to do with evaporation rates. You need to use TC treated plates for your cell culture.
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