I would like to develop a co-culture modle using a airyway epithelial cell and a bronchial fibroblast. I have read some references suggesting that I should seed the epithelial cell in the upper chamber and seed the fibroblast in the bottom chamer. And the proper pore size is 0.4 or 1.0 micrometer. The most frequently chosen material is PET. I know that the air-liquid interface could be creat if I seed the epithelial cell in the upper chamber. But I am confused that is the 0.4 or 1.0 micrometer PET membrane semi-permeable? How can I make a decision that which cell could be seeded in the upper chamber and which could be seeded in the bottom chamber?
The orientation of the cell types is independent of the semi-permeable support pore size. The epithelia are on the filter insert. The fibroblasts are either in the plastic in the receiving well or seeded onto the underside of the filter. Seeding on the plastic will allow examination of soluble factors that exert an effect. Seeding on the filter underside can possibly allow for effects that are exerted by cell-cell contact assuming that cell growth processes can pass through the filter pore. Likewise, a larger pore size can sometimes result in the migration of cells from the upper to the underside of the filter resulting in double epithelial monolayers. Another consideration of pore size is related to what you'll do with these co-cultures. The smaller pore size could potentially impede transfer of larger molecules.
In general, in the case of airways, epithelial cells are exposed to air, and fibroblasts are located inside the basement membrane. Therefore, it is appropriate to seed epithelial cells in the upper chamber for air-exposed culture of epithelial cells. The seeding location of the fibroblasts is up to your choice, but you can usually place them at the bottom of the chamber (the same goes for endothelial cells).
For the permeable membrane included in the cell culture insert, there are various materials such as PET and PC. PET is generally more transparent than PC, so it will be easier to observe cells. In addition, a smaller pore size is one of the variables that should be well considered because it can create a more stringent air-exposed culture environment.
I have performed quite a lot of coculture experiments to obtain corneal epithelial cell products for regenerative medicine. One of the reason why epithelial cells should be put in upper chamber, is to get the multiple layered products after air lift. The soft bottom of the upper well could by removed thus obtain the multi-layered epithelial cell sheets for transplantation
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