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Cause of Death for Cell Culture?


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#1 occamsrazorwit

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Posted 15 July 2011 - 11:14 AM

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?

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#2 Greg S

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Posted 15 July 2011 - 06:37 PM

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?

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?



#3 occamsrazorwit

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Posted 15 July 2011 - 08:03 PM

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.

#4 occamsrazorwit

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Posted 21 July 2011 - 07:22 AM

Also, this is what the cells in the culture look like.

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#5 casandra

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Posted 21 July 2011 - 12:54 PM


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 itís contamination or a bad cell lineÖ..you can try paired seeding (i.e. if you havenít done this already).Ö.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...
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#6 occamsrazorwit

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Posted 25 July 2011 - 09:51 PM

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.




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