I got LNCaP-FGC cell from NCCS-Pune. When it arrived it was in good health but after 2 passages it started turning bad, lots of black dots appeared beside the cells and eventually cells died after a week. In NCCS guideline they advice using a gelatin coated flask for growing these kind of cells as they grow very slow and adhere loosely. Getting a gelatin coated flask, plates and 96-well plate is difficult so i wanted to know if its possible to grow without it. Please help me with the answer.
In my experience gelatin coated flasks are not necessary (we got out LNCaP-FGC from ATCC). However, I made the experience that LNCaP grow better or worse on different culture plates from different manufacturers. If possible, try this out. Also don't split them too much (I usually split twice per week at 1:3 or 1:4). They like to have some company and grow more slowly if they are all alone!
We also don't use gelatin coated flasks. LNCaP indeed grow slowly and can be delicate, cells can be clumpy, and not as adherent as we might like (we tend to refer to them as a 'semi adherent' cell line). For us, we make sure not to under- or over-trypsinise as this can cause issues with this cell line. Under-trypsinizing in particular can lead to clumps. Following neutralisation of the trypsin, we pipette up and down at least 25 times, which sounds like a lot, I know. If the cultures are already clumpy, and you're trying to rescue them, I'd pipette up and down double or triple that many times... Again, sounds weird I know. The trick is to pipette up and down many times to avoid or disperse clumps, but to do so gently, as the cell line is also relatively delicate. We also passage twice per week, and we also tend to passage at around 1:3. I hope this helps!
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