I'm using poly-L-ornithine solution and fibronectin to coat plates for LUHMES cells. Since poly-L-ornithine is a little expensive, can I re-use the solution after the coating to coat more plates later? Thanks a lot in advance!
Absolutely! We routinely save and re-use for several times. Just keep a note of how many times on the bottle and open a fresh one every so often. There's no hard and fast rule to it, sometimes for 10+ uses and the cells still adhere fine. It's the laminin or anything extra you may coat with on top that is more sensitive than the poly-ornithine. (I culture mouse and human ESC/iPSCs-neural derivatives on this, they are super sensitive to changes in environment). Good luck
That`s no Problem, I used it several times too. And I did not have any Problems with growing the cells
Good luck!!!;-)
Thanks for starting this thread! I started doing this and it's working fine for me too!
I am also using LUHMEs and I coat with poly-L-ornithine overnight followed by a three hour incubation with fibronectin.
Question: can both be used simultaneously?
We incubate poly-L-ornithine and fibronectin simultaneously for at least 3 hrs in the incubator but preferably overnight.
In addition to PLO and fibronectin, has anyone tried coating the plates with anything extra like PDL, laminin or matrigel? When we grow the LUHMES neurons on glass coverslips that lift off very easy.
Jonathan, we culture mESC and differentiate them into neurons with a poli ornithine and laminin coating using coverslips and works just fine :)
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