See the Cell Migration/Invasion Overview for background information. The example below demonstrates the use of ACEA’s cell invasion and migration plate (CIM-Plate®) and the xCELLigence® RTCA DP system to study transmigration, with specific focus on cancer spheroid invasion.
Application Highlight: Studying Ovarian Cancer Spheroid Invasion of Mesothelial Cells
Once ovarian cancer cells shed into the peritoneal fluid, they aggregate and then bind to and invade through the peritoneal lining to establish secondary tumors. The complexity of this metastatic process makes it difficult to study, particularly in a quantitative manner. In the example highlighted in Figure 1, a model of the peritoneal lining, including a basement membrane matrix covered by a mesothelial cell monolayer, was established within the upper chambers of a CIM-Plate. The chemotaxis-fueled invasion of cancer spheroids through these layers was then quantitatively monitored in real-time using the xCELLigence RTCA DP system.
Figure 1 Real-Time Transmigration Assay. (A) Assay schematic. To mimic the microenvironment that metastasizing ovarian cancer cells encounter when invading through the peritoneal lining, the upper chambers of a CIM-Plate were first coated with a layer of Matrigel® followed by a monolayer of LP9 mesothelial cells. Finally, ovarian cancer spheroids were added on top. (B) Phase contrast microscopy image of ovarian cancer spheroids on top of an LP9 cell monolayer. (C) Quantitative analysis of cancer spheroid invasion capacity assessed 24 hours after plating. Media in the lower chamber of the CIM-Plate was either serum free (SFM) or was supplemented with FBS to drive chemotaxis. The capacity of LP9 cells to invade through matrigel (i.e. in the absence of cancer spheroids) is shown as a negative control. (D) Same as (C), except the real-time impedance readings are plotted continuously for 35 hours after spheroid addition. Figures adapted from J Vis Exp. 2014 May 20;(87).
Though ovarian cancer is the focus of the above study, the same methodology can be used to study other metastatic processes, including breast cancer metastasis or the extravasion of cancer cells through the endothelial layer. It can also be used to study normal cell invasion, such as trophoblast invasion during embryo implantation.
Key Benefits of xCELLigence for Transmigration Studies:
- Quantitative monitoring of cell invasion in real-time.
- Label-free assay requires no fixation, staining or any other sample processing, dramatically reducing hands-on time.
- Easy quantification of the kinetics of cell invasion.
- Rapid optimization of cell density and extracellular matrix density conditions.
- Non-invasive nature of the assay allows for cells on either side of the microporous membrane to be analyzed by complementary techniques (imaging, RT-PCR, etc.)
To see examples of how the xCELLigence RTCA DP system and CIM-Plate have been used for other types of cell migration/invasion studies, click the below links:
- Chemotactic Migration
- Wound Healing
Featured xCELLigence RTCA System for Cell Migration and Invasion
|xCELLigence RTCA DP|
Visit the xCELLigence RTCA DP product page for additional information