Spheroid Confrontation Assay

The research of the interaction and invasion behavior plays a crucial role in
the studies of the mechanisms of cell invasion which is significant in malignant
metastasis of cancer. Both the determination of the interaction and invasion
behavior can be achieved with the spheroid confrontation assay. And this
assay can monitor cell mobility in a three-dimensional model in vitro which is a
great advantage compared to other invasion assays based on the two-
dimensional modal.
In this approach, two performed spheroids derived from two different cell
types, one is of invasive character and the other is noninvasive type. Both of
them are cultivated side by side, which eventually commence to fuse. Both of
the two different cell types are fluorescently labeled with two different dyes
and spheroids of these cells are formed in a medium with proper
concentration of methycellulose. Therefore, we can easily monitored cells
either infiltrating the opposing spheroids as single cell or as collectively
exhibiting a non-invasive phenotype. In the non-invasion case, a distinct
margin is formed at the interface between the two cell types. The invading
cells can be quantified by either confocal imaging system or
immunohistochemistry analysis including paraffin embedding and tissue
section. If the cells express distinct markers, cells can easily be differentiated
immunohistochemically.
 Figure 1. The

experimental results of Spheroid Confrontation Assay

The spheroid confrontation assay makes it possible to study cell invasion in

3D model, which is closely reflecting the vivo situation especially in case of a
carcinoma is invading a certain tissue or organ. It also can be used to study
the cell invasion under the situation cell-cell interaction, which could mimic the
situation that tumor cells invade the normal tissue. Cell invasion studies with
spheroid confrontation assay lead a new insight to the mechanisms of cell
invasion in vivo. The advanced confocal imaging system and experienced
experts at Creative Proteomics guarantee a reliable and reporducible result of
the spheroid confrontation assay.