Hi Sílvia :)

That's normal behaviour for almost any type of cell sheets I know of. You have two options for that. You can use a carrier to transport one cell sheet on top of other and then wait for the sheets to bind. This requires optimization but while attached to the carrier the cell sheet won't shrink. Okano's group have developed a gelatin-based manipulator that acts as a carrier and helps in the stacking. You can build your own. The other option is to use ROCK inhibitor in the culture before detaching. I remember seeing this in a presentation of someone from Okano's group. It apparently avoids CS shrinkage.

Hope it helps. 

R

Hi Silvia,

I was working at Okano's group though I have not tried to stuck EC sheets by myself.

Roger, The ROCK inhibitor thing might be my topic focusing on cytoskeletal dynamics using oral mucosal epithelial cell sheet. I think it should work, but it can be a bit tricky to start with as a stacking method. I'm also not sure about ROCK inhibitor's side effects on other types of cells.

Basically we are using gelatin-coated cell sheet manipulator (stamp) method to stuck EC sheet beautifully. There are several literatures you can find like this one. 

http://onlinelibrary.wiley.com/store/10.1111/j.1365-2796.2009.02185.x/asset/j.1365-2796.2009.02185.x.pdf?v=1&t=in5osx0j&s=830cc21ad87f8bc1b74dd2c01fc9486f8c6f2ae0&systemMessage=Subscribe+and+renew+is+currently+unavailable+online.+Please+contact+customer+care+to+place+an+order%3A++http%3A%2F%2Folabout.wiley.com%2FWileyCDA%2FSection%2Fid-397203.html++.Apologies+for+the+inconvenience.

Hope this will help.

Makoto