I'm working with prostate cancer from dogs and is not a very common disease. I have some frozen tumor and I have seen few protocols to generate cell culture from frozen tissue. Is it really possible to generate cell line from frozen tissue?How do you establish a cell line from the frozen tissue sample?
Indeed a nice thought, was thinking in same lines, we have tries on conditionally frozen sample human tissues. However, we could not succeed much the cells isolated were dead and could not grow at all. So in short it is difficult.
I think it's possible, I'm working on Mesenchymal Stem Cell from umbilical cord matrix. How do you cryopreserve the tissue? some journal I've read say that it depend on how you cryopreserve the tissue.
Hi Carlos!
In short, it's tricky, which doesn't mean impossible. How did you preserved your samples? If the tissue was snap frozen in liquid nitrogen then it is unlikely. But if it has been properly cryopreserved using DMSO then look at this thread... http://www.protocol-online.org/biology-forums/posts/15958.html
Hope it helps. Good luck!
Hi Carlos,
important point is cell line of what cell type would you like to generate?
Theoretically, different cell types (epithelial cells, endothelial cells, fibroblasts etc) will have different sensitivity to the cryopreservation (and will further depend on the method used for it).
So, some cell types could (theoretically) survive and you will be able to rescue them and put in a culture. But some cell types will die - and you will get non physiological "primary culture".
Another moment - even if you will rescue some individual cells and get a culture, their morphology/metabolism could be distorted by the cold-induced stress. As the result, it will be difficult to differentiate prostate cancer mechanisms from stress-induced changes.
In this situation, it could be complicated to use the obtained "cell culture" for sensitive experiments.
May be it is more reliable to wait for a fresh material and be sure in the quality of your cell culture model.
Hi Kumaravelu, Stefan and Norbert,
Thank you very much for your answers. My samples were cryopreserved using DMSO.
Norbert, Thank tou very much for the protocol. It helped me a lot.
Hi Elvira,
I would like to generate culture with epithelial cells. Neoplastic epithelial cells. I had not thought about alterations in cell morphology/metabolism. It 's a good point. Thank you so much for the advice. I will think about it and maybe I will wait for a fresh tissue.
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