PBS and Cells - How long can cells sit in PBS? (Jul/11/2006 )
How long do you think a cell could sit in 1X PBS without it having an adverse effect? Would it depend on the cell type (in this case Hepa 1-6)? I'm trying to come up with the best way to prepare cells for surgical implantation and one suggestion was to dilute them in PBS. I just don't want them to end up sitting in PBS for too long and die. The other suggestion was just to dilute the cells in media.
I might leave the cells in the medium and right before implanting the cells, spin it and resuspend them in PBS.
You need to minimize shocking the cells. This includes osmotic shock, chemical shock and whatever physical parameters are involved..temperature?
Suspending your cells in PBS exposes them to the right pH for subsequent implantation but I'd worry about the phosphate and certainly the sodium concentrations. I'd check the osmolality of PBS. (needs to be around 295mosmols).
Would it be possible to resuspend the cells in the patients own serum before implantation? They must have had blood samples taken for Us&Es, LFTs and maybe tumour markers so the (hospital) lab may be able to dig out a smaple for you.
Suspending your cells in PBS exposes them to the right pH for subsequent implantation but I'd worry about the phosphate and certainly the sodium concentrations. I'd check the osmolality of PBS. (needs to be around 295mosmols).
Would it be possible to resuspend the cells in the patients own serum before implantation? They must have had blood samples taken for Us&Es, LFTs and maybe tumour markers so the (hospital) lab may be able to dig out a smaple for you.
The patient would be a mouse (and as far as I know they haven't taken blood from it) ... so I'm not too sure whether or not suspending the cells in it's serum would be possible ... but it is a nice idea and I may suggest it. I will be keeping the cells in the PBS at 37C until time of surgery and I will definately check out the osmaolality of the PBS. I'm preparing the cells for one of the PhD students in my lab and this is his first experience with implanting cells. I've prepared glioma cells before and kept them in a mixture of media and 1% agarose for implantation and this seems to work. However they don't want to use agarose. I suppose for the first few surgeries all of this will be trial and error.
I may try and experiment and see what the viability of my cells will be in PBS over time.
medium is used very frequently and is better if you want to wait for long time.
Do you know if antibiotics in the medium would affect anything?
u could use medium without antibiotics. For a short period of time, it shouldn't matter.