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

After centrifuge in serum-free media, forming cell sheet, instead of pellet in t - (Jul/11/2014 )


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?




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?




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?