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Keratinocyte Morphology in Monolayer Culture


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#1 NazGoole

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Posted 19 November 2013 - 11:12 AM

Hi everyone,

 

I am currently working with human keratinocytes. I seed them into Flasks and let them grow until I can use them for my experiments.

Some of the cultures are doing very well so far, but others at least seem a bit difficult.

They do not all have these "cobblestone" aspects. In some flasks all of the keratinocytes are just small and round, although attached (so I guess they must still be alive) and covering almost the entire flask.

As I haven't found any information about this "behaviour" of the keratinocytes, I wanted to ask you guys if per chance one of you can tell me what the hell is going on with these keratinocytes.

Perhaps someone has or knows about some kind of atlas where u can browse through all possible shapes of these cells?

 

Sincerely,

NazGoole



#2 roxx_gurl

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Posted 19 November 2013 - 07:15 PM

Hi

 

What is the origin of your keratinocytes? Is it from skin? and is it primary or cell lines? Are you using enzymatic or explant technique? To be honest keratinocytes is pain in the ass, and you need to be patient. If you are using explant technique, the round cell might be contaminated cell. Keratinocytes is specifically has the characteristic of cobblestone when they attached and grow. That is all. If regular round shape, I am afraid that is not keratinocytes. Anyway what culture media you are using now?

 

Hope it helpsrolleyes.gif



#3 CellApplicationsInc

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Posted 20 November 2013 - 10:13 AM

Hi NazGoole,

 

this video of primary Human Epidermal Keratinocytes that I just posted on our blog might be helpful. 

http://bit.ly/1aRGyHk

 

Good luck!



#4 NazGoole

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Posted 21 November 2013 - 02:19 PM

Hey thanks for your replies smile.png

well the Keras are from patients, and I extracted them with enzymes (Trypsin etc.) so they are primary. Now many of them have started to grow and form the typical spines but still no islands. Looks like I might have some fibroblasts in the flasks.........

Thanks for the video, thats really helpfull too I just bookmarked it, so please do not delete it wink.png
My cells looked like those white dots in the beginning of the video but it takes a bit longer for them to differentiate in overall. I barely ever get the cobblestone aspect that quickly.... sad.png
But I use serum-free medium that might complicate things.

Thanks a lot! smile.png


Edited by NazGoole, 21 November 2013 - 02:28 PM.





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