After I have trypsinized mammalian cells and removed them from a flask, can I reuse that flask to seed the same culture? This is assuming that I used good aseptic technique. I mainly want to do this when a culture becomes to confluent and I just want to reduce the number of cells.
I am doing it constantly with different cancer cells. In fact they even grow better in the used container, than in fresh one. I am using the same flask even for two-three weeks. It greatly reduces the cost of cell culture. And with good aseptic technique (as it was mentioned before) there would be no problems.
In my experience, it depends on the cell line. I've seen some cells that don't mind and will sit down just fine on the original plate. Others, not so much. I would test it in parallel plates. Good luck!
If you perform good aseptic technique and proper rinsing of the vessel during your culture, I don't see why not. Some researchers might be against this as some leftover metabolites and/or enzymes from the previous culture (especially with mammalian cell culture) might adhere to the culture vessel's surface, but in my experience, I have not had major problems when I reuse my culture vessels for the purpose of maintaining my cells. The practice in our lab is to reuse the same culture vessel not more than 2x before switching to a new one, just to be safe from possible contaminants which may affect the growth of the culture.
I am doing it constantly with different cancer cells. In fact they even grow better in the used container, than in fresh one. I am using the same flask even for two-three weeks. It greatly reduces the cost of cell culture. And with good aseptic technique (as it was mentioned before) there would be no problems.
The only problem I can see with flask reuse is that the "cell culture treatment" that the manufacturer adds to enable the cells to attach and grow may degrade over culture time. However, since there are so many undefined variables in this, just testing in parallel cultures should suffice. I usually split cultures (and after cell counts) reintroduce an aliquot int the original flask(s) and use the rest to seed new flasks, particularly when expanding cell lines.
Actually it depends on various factors. But, if you are working with the same cell line, same treatment it will be okay. But, if your cells get spoiled, please don't reuse it.
Hi
How do you perform aseptic techniques to reuse flasks and
petri dishes?
If you are working with Petri dishes, the best way to reuse them is to use glass Petri dishes and wash, thoroughly rinse, and autoclave them after each use. They can be used for years. The old cleaning methods involved boiling in a solution that when rinsed afterwards left a new microscopically thin layer of new glass on the surface. The newer methods involve soaking in Microclean and rinsing 10 times in running tap water, followed by a distilled water rinse, air dry and autoclave..
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