Development of an Insert Co-culture System of Two Cellular Types in the Absence of Cell-Cell Contact

The overall goal of this procedure is to create a cellular co-culture system of two cell types using inserts with a permeable membrane, thus allowing the diffusion of secreted soluble factors. This is accomplished with a series of steps that finely placed inserts containing one cell type in a multi-welled tissue culture plate containing a second cell type. We start my seeding cell type number one in inserts, and seeding cell type number two in a multi-well tissue culture plate.

The second step is to transfer the inserts into the wells of the plate containing cell type number two. Ultimately, this gives the ability to harvest the two cell types as well as the respective supernatants. Therefore, it is possible to evaluate the effect of secreted soluble factors on the cell type of interest.