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What happened to my cell? (Colony formation)


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#1 Clover

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Posted 20 June 2014 - 12:37 PM

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. :(

 

Screen Shot 2014-06-20 at 3.28.30 PM.png

 

Clover



#2 bob1

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Posted 20 June 2014 - 01:39 PM

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



#3 Clover

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Posted 25 June 2014 - 03:20 PM

mt8pc3.jpg

 

How about this one? Apoptosis? 

 

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

 

2vdle8i.png



#4 bob1

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Posted 25 June 2014 - 03:43 PM

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?



#5 Tabaluga

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Posted 25 June 2014 - 10:24 PM

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

Il dort. Quoique le sort fût pour lui bien étrange,
Il vivait. Il mourut quand il n'eut plus son ange;
La chose simplement d'elle-même arriva,
Comme la nuit se fait lorsque le jour s'en va.

 


#6 Clover

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Posted 26 June 2014 - 08:47 AM

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.)



#7 Clover

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Posted 26 June 2014 - 08:49 AM

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?



#8 Tabaluga

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Posted 27 June 2014 - 03:35 AM

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.


Il dort. Quoique le sort fût pour lui bien étrange,
Il vivait. Il mourut quand il n'eut plus son ange;
La chose simplement d'elle-même arriva,
Comme la nuit se fait lorsque le jour s'en va.

 





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