Hi,
I have extensive 2D and 3D spheroid cell culture experience and I am considering a multi-cellular culture whereby I layer cell types on top of each other, for example stromal fibroblasts followed by epithelial cells.
Is it as simple as timing the addition of the second layer (the top) of cells addition so that both layers become confluent at a similar time, providing I use the correct mix of media and supplements for both cell types?
Or is there a little more to it such as an ECM coating of say fibronectin to stick the second layer to the first layer?
Many Thanks,
Ethan
Dear Ethan
How about using the technique described in the paper attached below?
The investigators have used the concept of stacks, a reconfigurable multilayer suspended microfluidic system. This reconfigurable stack system will allow researchers to study complex multicellular interactions by offering the opportunity to independently prepare each microenvironment and then combine them in any desired sequence and combination.
Please refer to the paper for more information.
Article Reconfigurable open microfluidics for studying the spatiotem...
Hope it is helpful!
Best.
If you don't want to use robot -printed matrices or microfluidics, the old fashioned way is to seed the fibroblasts in a collagen coating , then plate the epithelial cells onto the collagen. An example:
Article A 3D epithelial–mesenchymal co-culture model of human bronch...
If you are interested in diffusible factor effects, cells can be seeded on opposite sides of cell culture inserts. https://www.sigmaaldrich.com/US/en/technical-documents/technical-article/cell-culture-and-cell-culture-analysis/mammalian-cell-culture/basic-indirect-co-culture
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