Protocol Online logo
Top : New Forum Archives (2009-): : Tissue and Cell Culture

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. :(

 

Attached Image

 

Clover

-Clover-

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

-bob1-

mt8pc3.jpg

 

How about this one? Apoptosis? 

 

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

 

2vdle8i.png

-Clover-

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?

-bob1-

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

-Tabaluga-

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? 

 

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

-Clover-

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. 

 

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?

-Clover-

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.

-Tabaluga-