What happened to my cell? (Colony formation) - (Jun/20/2014 )

Hi,

I am growing cell colonies in agarose gel. And in this picture, it looks like my cell opened a window? What happened?

BTW, this is U251, glioma cell. It has been cultured within the gel for five days without growing. :(

Clover

It is quite an atypical looking cell, but it looks to me like it is senescent, where you often get vacuoles opening up.

How about this one? Apoptosis? 

I have a photo looks exactly like apoptosis, but they two does not look alike.

It's very hard to say - what magnification are you using?  Is it a single cell or a colony?  what conditions are you culturing the cells under?

My very first impression of the second photo you posted is that they are indeed dead ( also did colony formation with glioma cells.)

Tabaluga, I'm really glad you came. Cause I had been struggling to find a way to make my cells growing in gel.

I tried 0.7% and 0.6%, but it does not change much. Could you please share some of your culturing conditions? 

Tabaluga on Thu Jun 26 06:24:22 2014 said:

My very first impression of the second photo you posted is that they are indeed dead ( also did colony formation with glioma cells.)

It's a single cell, *20 I think. Just normal DMEM(With L-Glutamine, 4.5g/L Glucose and Sodium Pyruvate; without Sodium Bicarbonate)+0.6-7% gel. 

bob1 on Wed Jun 25 23:43:33 2014 said:

It's very hard to say - what magnification are you using?  Is it a single cell or a colony?  what conditions are you culturing the cells under?

Sorry to have to disappoint you Clover, but I actually did it with glioma cells that were already growing naturally in the spheroid form without agarose, so it's not the same as in your situation. Also I did not seed single cells but 1000 cells.

I used 0,67 % low melting agarose.