Dear All,
For the environmental purposes and also saving money, can cell culture flasks be reused?
I am using an adherent cell line. Whether T75or T175.
Thanks!
I have successfully re-used flasks multiple times.
You just need to be very careful to avoid contamination.
Hello Farbod Bahreini
Nice to hear that you care for the environment!
Yes, you may reuse T75 or T175 flasks, but at the same time I would also like to share the following information.
For suspension cells, there is no problem re-using the flask many times (at least 10-12 times). But with continuous use, you will come across some type of residue on the plastic surface . So just be on a look out for such deposits. After a few cycles, if you notice that your flask is now not suitable for re-use, you may discard the flask.
On the other hand, for adherent cultures, it is recommended that the flask be re-used 3-4 times. This is because tissue culture treated flask are given special treatment to ensure consistent growth surface which may degrade over continuous re-use. Also, constant re-use of tissue culture flask may result in the deposition of adhesive proteins by cells of previous passages on the surface of the flask resulting in cells with altered growth and metabolism. You will also observe that adherent cells repeatedly grown on such surfaces become difficult to trypsinize in the later cycles.
So, if you are using adherent cell line, you may re-use T75 or T175 flasks 3-4 times. I re-use T-75 flasks for adherent cultures 3 times without any negative effect on cell growth.
Best.
Hi Farbod Bahreini
I reuse culture flask all the time. It's better to use a new one to avoid and minimize chances of contamination but if you are careful enough this can be done on regular bases. I do it all the time. In fact I have reused even culture plates like 6-well plates. You can easily reuse these items as long as you take full precautions to avoid the risk of contamination.
Reuse is very useful in reducing research costs, but you always need to have an alternative cell culture in case of contamination.
I have reused a few times culture flaks, indeed, it is possible after lots of decontamination (several washes and UV) but it is always risky so, the times i've reused is only in urgency.
T175 flasks are commonly used for cell culture but are typically considered disposable. However, under certain conditions, they can be reused with proper cleaning and sterilization. Factors to consider include the material and condition of the flask, the risk of contamination, the cleaning and sterilization process, and conducting quality control tests. Polystyrene flasks in good condition can be suitable for reuse if thoroughly cleaned and sterilized. However, complete removal of all contaminants can be challenging, so careful cleaning and quality control are essential. The number of effective reuses may vary depending on factors such as culture conditions and cell types.
The T175 flasks, which are often utilized in cell culture, are usually regarded as single-use items. However, its reuse when working with the same cell line presents some advantages, such as the presence of an extracellular matrix consolidated by the previous culture. This is an enriched network of proteins and other molecules that help the cells adhere to the flask.
Short answer: yes.
Long answer: depends. Some cell lines are very dependant on extracellular matrix and would thrive in a flask that have alredy been colonized by cells and have built an extracellular matrix. Also, cells are prone to relese growth factors that stimulate proliferation, so that can be beneficial. Other cell lines may release DAMPs when reaching a high confluence and that may be left on the flask you are reusing and affect cell growth. Besides, you can eventually generate a trypsin resistance because the build up ECM and cells that did not separat properly form the old flask. Other factors may also be implicated, like differentiation, senescence, transformation, etc. I think you should consider if the benefits outweight the costs in your case.
here is a nice paper you can read on DAMPs: Article Damage-associated molecular patterns (DAMPs) related to immu...
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