Cellular Invasion

The ability of malignant tumor cells to invade normal surrounding tissue contributes in large part to the morbidity and mortality of cancers. Cell invasion requires several distinct cellular functions including adhesion, motility, detachment, and extracellular matrix proteolysis.

Cell invasion assays have become a critical technique in screening compounds that can inhibit metastasis or promote angiogenesis.

Technology Overview

Cellular Invasion is done using Oris kits. In this assay a rubber cell stopper is used which can be fixed on the plate. This stopper does not allow the cells to adhere in the center of the well (Figure 1). This inhibition of the cell growth acts as a detection zone. The cells are overlaid will Collagen Type I and incubated. Cells invade and cover the detection zone. After a fixed time point the cells are stained with a fluorescent dye and read on a plate reader.

Figure 1. HT-1080 Cell Invasion Microscopic Data Ref. Oris Technologies

Service Design

Clients can submit plant extracts, synthetic compounds, peptides, formulations etc for invasion testing using MTT assay. Invasion can be tested on various human normal and cancer cell lines. More details about the cell lines can be obtained from DL cell line directory or by contacting us. We provide customized assay design. Customers can select the parameters like

• Drug concentration to be tested
• Time of incubation of the drug with cells
• Duplication of well
• Internal standards
• Cell line

End Point Detection

Clients can screen huge libraries of test compounds at a fixed single concentration to find the interesting inhibitors or activators of invasion called “Hits”. Such an assay format is very economic and fast to find interesting molecules which can be further profiled for IC50 determination. This assay is very helpful for chemists to evaluate compounds with diverse chemical structures and find the most interesting structures to start with.

IC50 Determination

IC50 determination can be performed to further evaluate the hits. Compounds can be tested at 5-8 concentrations and can be evaluated for cellular invasion at a fixed time point. Percent closure of the detection zone can be calculated and plotted against concentration to calculate IC50. This provides more information about the activity of the test sample at different concentrations i.e. dose- response curve. Samples can be cross compared using IC50 values. This information is very helpful for the chemists to do the Structural Activity Relationship (SAR) for further modifications

Kinetic Detection

Detection zone closure can also be studied at various time points till a complete closure is obtained. Such studies are important to compare cell lines, compare cells over expressing a particular gene or effect of test substance with time.