There are several methods to measure cell adhesion which are usually based on washing or shaking to remove non-adherent cells. Meanwhile, cells are quantified only when they are in contact with the adherent surface and they are separated. In this case, the anti-adhesion effect of some components like hormones and cytokines might be away during this frame time. Here, Creative Proteomics develops a quantification method which can separated non-adherent cells and only quantify vital cells with colorimetric assay. This method can be used especially when the “anti-adhesion” effect is present only for a shot time such as peptides and cytokines that only provide a trap for non-adherent cells in a way that they can’t touch again the adherent surface. This method facilitates fast and reliable measurement of cell adhesion in multiwall format for screening assays.
This assay is designed to quantify living cells and can be applied as broad as researchers need. Initially, the adherent cells are seeded in one 96-well plant and incubate them until there is a confluent layer. Then, the medium of the adherent cells is removed and add new culture medium together with the cell suspension which will be tested and incubate the plate for a reasonable time in order to let the cells seed and make contact with the adherent cells. A new 96-well plate in the laminar flow hood is placed. Take the plate which incubate the adherent cells with the cover as it is (upside down) and place it in the laminar flow hood (also upside down). Finally, colorimetric reaction and measurement of non-adherent cells will be conducted.
Figure 1: The experimental process of Cell-Cell Adhesion Assay
This new adhesion technique has several advantages comparing with other methods:
1. Reduction of the time period during the "test" and the "processing" of results.
2. The experiments can be done in a larger scale.
3. Several conditions can be tested at the same time.
4. Quantification of vital cells.
5. It is possible to separate the cells which were non-adherent and use them for further examination.
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