I have been trying for months to grow kidney cells from rat kidneys, I tried several methods, serial sieving, cutting and meshing, and using kidney tissue to obtain an outgrowth of cells. I always end up with what looks like cell debris but they keep increasing, which indicates that they are live cells, but they don't look like it. Any idea what is going on here? I have to say that these are mostly adherent and very hard to detach too, I used trypsin and trypLE... but some of them float too, just like any cells. Any pointers to what this could be is really appreciated, is this a contamination of some kind?
Dear Amal ... we are following an established protocol thta worked perfectly fine in our hand for primary tubular cells (https://www.hindawi.com/journals/omcl/2008/471575/).
I am not sure what kind of medium you used to grow these cells, but I would suggest adding epidermal growth factor to help the tubular cells to attach and grow. You may also try coating your plates with 1% Collagen-1 to help the attachment of the cells.
The image is not clear, but it is normal to have cell debris with the isolated cells, but most importantly you should notice cell proliferation by the second or third day of culture. The protocol provided is detailed and contains the supplements that should be added to the culture medium (we use DMEM/F12). I hope this answer is somehow helpful.