I have mouse cortical astrocytes which have already been passaged twice and are in healthy condition but want to passage them again for my experiment. Is it safe to do ? And do I need to use the precoated plates for culture of astrocytes. Any kind of suggestion is most welcome.
Thanks
Hi Ravinder,
in our hands astrocytes are safe to passage many times (8-10 times even) without affecting viability. I agree with Oleg that astrocytes are reactive even in short term cultures. We use them to study their expression of glial scar molecules (certainly a reactive phenotype) and find them to be quite reactive even in control conditions on the first passage.
Hi Ravinder,
We manage to use them up to 10 passages or less. At some point, you can easily see that they don't attach well and don't proliferate as well as before. However, I have to say that nothing is better than fresh primary astrocyte cultures so that you avoid using trypsin...
Sighild
Thanks for all the answers. I too am wondering. In our lab, people used to do only freshly isolated astrocytes. Now, I have used between 3-5 passages. It seems to be working well but I wouldn't risk to many passages. One good indicator of cell viability (crude way) is their adhering power as Michael pointed out.
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