Cause of Death for Cell Culture? - (Jul/15/2011 )
I've been culturing various cell lines and running experiments. Recently, I've been focusing on PC-3 (prostate cancer) cells in a PDMS (polydimethylsiloxane) device but these cancer cells keep dying on me. The culture that I used to seed cells into my PDMS device is very healthy, but, a day after seeding, these cells all die. Most of the cells have underwent apoptosis and the whole microscope view is full of tiny cell fragments (sorry for the crappy microscope pictures). The device is surrounded by PBS and sterilized by soaking in alcohol.
Can anyone identify the problem for me?
Not totally sure, but 2 things spring to mind:
1) They might not be thrilled with being surrounded by PBS if they are cultured overnight.
2) Maybe there is some kind of toxic interaction wtih the PDMS? Are PC-3 cells normally adherent? Perhaps they're not thrilled with touching PDMS vs normal PS/PP in cell-culture dishes?
occamsrazorwit on Fri Jul 15 19:14:40 2011 said:
I've been culturing various cell lines and running experiments. Recently, I've been focusing on PC-3 (prostate cancer) cells in a PDMS (polydimethylsiloxane) device but these cancer cells keep dying on me. The culture that I used to seed cells into my PDMS device is very healthy, but, a day after seeding, these cells all die. Most of the cells have underwent apoptosis and the whole microscope view is full of tiny cell fragments (sorry for the crappy microscope pictures). The device is surrounded by PBS and sterilized by soaking in alcohol.
Can anyone identify the problem for me?
Greg S on Sat Jul 16 02:37:59 2011 said:
Not totally sure, but 2 things spring to mind:
1) They might not be thrilled with being surrounded by PBS if they are cultured overnight.
2) Maybe there is some kind of toxic interaction wtih the PDMS? Are PC-3 cells normally adherent? Perhaps they're not thrilled with touching PDMS vs normal PS/PP in cell-culture dishes?
In previous similar experiments, I have surrounded the device with PBS with no ill effects. The PBS does not come into contact with the cells and mainly serves to stop evaporation of the media. Also, the PDMS is used because it doesn't interact biologically. PC-3 cells normally adhere in this device.
In response to the second post, the pH is normal for these cells. Could it be due to the alcohol presoak of the devices, contamination, or a bad cell line? All of the alcohol should have dried off of the device before I used it.
Also, this is what the cells in the culture look like.
occamsrazorwit on Sat Jul 16 04:03:37 2011 said:
Greg S on Sat Jul 16 02:37:59 2011 said:
Not totally sure, but 2 things spring to mind:
1) They might not be thrilled with being surrounded by PBS if they are cultured overnight.
2) Maybe there is some kind of toxic interaction wtih the PDMS? Are PC-3 cells normally adherent? Perhaps they're not thrilled with touching PDMS vs normal PS/PP in cell-culture dishes?
In previous similar experiments, I have surrounded the device with PBS with no ill effects. The PBS does not come into contact with the cells and mainly serves to stop evaporation of the media. Also, the PDMS is used because it doesn't interact biologically. PC-3 cells normally adhere in this device.
In response to the second post, the pH is normal for these cells. Could it be due to the alcohol presoak of the devices, contamination, or a bad cell line? All of the alcohol should have dried off of the device before I used it.
hey orw,
if you're worried about residual ethanol killing off your cells, couldn't you "wash" the device first with sterile medium in the hood before seeding your cells? and if it’s contamination or a bad cell line…..you can try paired seeding (i.e. if you haven’t done this already).….use a common stock and grow cells in this device and in parallel, just in the regular tissue culture plate or dish and see what happens....at least it will give you clues if there's something wrong with your cells...
Thanks everyone! I have no idea what caused it and followed all of your suggestions. Strangely, the problem seems to have disappeared, so I guess I'll let it rest for now.