5 replies to this topic

[EvilTwin]

I've encountered a number of strangely looking cells in my HaCaT culture. About 5% of the cells look like they had a few bubbles inside. They are normal in size and shape but instead of grains and a hint of nucleus they seem to be almost completely filled with 3-20 large droplets of liquid (which irrefutably resemble vacuoles). The rest of the cells look healthy, I don't observe many dead cells in medium, nevertheless the growth is somehow inhibited.

I will check the cells for mycoplasma anyway, but such bubbles are not listed as a symptom of mycoplasma contamination. Does anyone have any idea what it could be?

Edited by EvilTwin, 17 April 2012 - 12:40 AM.

[bob1]

Senescent cells often get vacuoles in them.

[EvilTwin]

Thank you I guess it was because the poor condition of my cells. In case anyone has a similar problem: no infection developed, after subculturing 3 times my cells look very good. PCR didn't reveal mycoplasma contamination. It seems to me that the cells just didn't like freezing, because the phenomenon appeared right after getting them from the bank.

Edited by EvilTwin, 14 May 2012 - 12:44 AM.

[bob1]

Any given population of cells will have some senescent cells in it, so don't be too worried, unless you see a higher abundance than normal.

[h9494036]

Eviltwin:  Hacat cells also require a rather high cell density to grow.  If the density is too low, they do not like the environment and they start forming vacuoles.  Nothing wrong with the cells, I believe.

[bob1]

h9494036, on 10 October 2012 - 03:22 PM, said:

If the density is too low, they do not like the environment and they start forming vacuoles.

This is known as senescence and is irreversible for those cells that start it.