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After centrifuge in serum-free media, forming cell sheet, instead of pellet in t

serum free media cell culture centrifuge

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#1 roxx_gurl

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Posted 10 July 2014 - 09:29 PM

Hi

Has anyone encounter cell sheet across vertically in the tube, rather than normal cell pellet sink to the bottom after centrifugation ? How to get the supernatant out without disturbing the cell sheet?

 

Thanks



#2 bob1

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Posted 11 July 2014 - 01:19 AM

Are you sure it is cells?  This is quite unusual, but I could see it happening with fat cells. 

 

You could use a hypodermic needle and a syringe to minimize the damage to the layer?



#3 roxx_gurl

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Posted 11 July 2014 - 04:54 AM

Oh 

 

Yes. It forming cell floated instead of cell pellet sink to the bottom after centrifuge.  I have experienced this before using human gingival epithelium (normal) from Cell N Tec and using their culture media, I thought they must have differentiated due to confluency and maybe because of serum-free media. 

 

Btw, I tried to neutralized trypLE (can stop detachment by dilution)  with serum-containing media (maintenance culture using serum-free media), and after centrifuge, they formed cell pellet at the bottom like normal. 

 

I guess the cell pellet formation due to serum-containing media, somehow serum attach cell together and help to form cell pellet sink to the bottom,easy to manage

 

What do you think?







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