Cell Migration & Invasion Assays

The Boyden Chamber Assay
Cell Migration Assays
Cell Invasion Assays
Microfluidic Migration Device
In Vitro Scratch Assay
ECM Proteins
References

Metastasis is the cumulative result of multiple changes in tumor cells and their microenvironment that enables cellular migration and invasion into healthy host tissue. As proliferating neoplastic cells attempt to escape the primary tumor site, local cell adhesion and invasion of the surrounding tissue must occur¹. Prior to penetrating the blood vessel endothelium and gaining access to the blood stream, cancer cells must invade local tissues by degrading ECM protein components and ultimately, transverse the basement membrane². Once in circulation, these cells can form metastatic colonies at secondary locations.

Figure 1. Tumor metastasis is a multistep process involving cancer cell adhesion, cell migration and invasion into neighboring tissues and vasculature.

The Boyden Chamber Assay

The most widely accepted cell migration technique is the Boyden Chamber assay³. The classic transwell migration assay system uses a hollow plastic chamber, sealed at one end with a porous membrane. This chamber is suspended over a larger well which may contain medium and/or chemoattractants. Cells are placed inside the Chamber and allowed to migrate through the pores, to the other side of the membrane. Migratory cells are then stained and counted.

Figure 2. The Boyden Chamber Assay Protocol. Cells are allowed to migrate through a cell monolayer or ECM protein mixture which have been seeded onto a semi-permeable membrane cell culture insert with chemoattractants added below the membrane. Migrated cells can then be quantified by staining cells with DNA dyes such as Calcein-AM or CyQUANT GR Dyes

Cell Migration & Invasion Kits

Millipore’s QCM™ cell migration and invasion assays provide a quick and efficient system for the quantitative determination of various factors on cell migration, adhesion and invasion including screening of pharmacological agents, evaluation of integrins, chemoattractant or other adhesion receptors responsible for cell migration. The kits have provided researchers with consistent results for over a decade and have been published in over 4000 Publications.

Cell Migration Assays: Enables convenient and sensitive quantification of in vitro cell migration towards a chemical concentration gradient (chemotaxis) or ECM protein gradient (haptotaxis).

Cell Invasion Assays: Enables convenient and sensitive quantification of in vitro cell invasion through a basement membrane ECM protein or a layer of cells such as endothelial cells.

How to select the appropriate pore size for your cells:

• 3 μm pore size is appropriate for leukocyte or lymphocyte migration.
• 5 µm pore size is appropriate for a subset of fibroblast cells or cancer cells such as NIH-3T3 and MDA-MAB 231 cells. Also suitable for monocytes and macrophages.
• 8 μm pore size is appropriate for most cell types. This pore size supports optimal migration for most epithelial and fibroblast cells. Note - the 8 μm pore size is not appropriate for lymphocyte migration experiments.

Cell Migration Assays

Description	Pore Size	Plate Format	ECM Coating	Detection	No. of Tests	Product No.
Chemotaxis Cell Migration Assays	8 µm	24-well	None	Colorimetric	24	ECM508
		24-well		Fluorometric	24	ECM509
		96-well		Fluorometric	96	ECM510
	5 µm	24-well		Colorimetric	24	ECM506
		24-well		Fluorometric	24	ECM507
		96-well		Fluorometric	96	ECM512
	3µm	24-well		Colorimetric	24	ECM504
		24-well		Fluorometric	24	ECM505
		96-well		Fluorometric	96	ECM515
Haptotaxis Cell Migration Assays	8 µm	24-well	Fibronectin	Colorimetric	24	ECM580
		24-well	Collagen I	Fluorometric	24	ECM582
	5 µm	24-well	Laminin	Colorimetric	24	ECM220
		24-well		Fluorometric	24	ECM221

Cell Invasion Assays

Description	Pore Size	Plate Format	ECM Coating	Detection	No. of Tests	Product No.
Cell Invasion Assays	8 µm	24-well	ECMatrix™	Colorimetric	12	ECM550
		24-well		Colorimetric	24	ECM554
		96-well		Colorimetric	96	ECM555
		24-well	Collagen I	Colorimetric	24	ECM551
		24-well		Fluorometric	24	ECM552
		96-well		Fluorometric	96	ECM556
Endothelial Cell Migration Assays	3 µm	24-well	Fibronectin	Colorimetric	24	ECM200
		24-well		Fluorometric	24	ECM201
Endothelial Cell Invasion Assays		24-well	ECMatrix™	Colorimetric	24	ECM210
		24-well		Fluorometric	24	ECM211
Leukocyte Transendothelial Migration	3 µm	24-well	Fibronectin	Colorimetric	24	ECM557
Tumor Cell Transendothelial Migration	8 µm	24-well		Colorimetric	24	ECM558
QCM™ Invadopodia Gelatin Degradation Assay (Green)	NA	NA	FITC-Gelatin*	Fluorometric	32	ECM670
QCM™ Invadopodia Gelatin Degradation Assay (Red)			Cy3-Gelatin*	Fluorometric	32	ECM671

Microfluidic Migration Device

Millicell^® µ-Migration Assay Kit overcomes the limitations of traditional multiwell migration assays. The innovative design of the µ-Migration Slide promotes a stable diffusion-generated concentration gradient that is consistently linear and lasts for more than 48 hours. Made from a plastic with high optical qualities similar to those of glass, the µ-Migration Slide is specially designed for video microscopy assays. At specific time intervals, images of the observation area can be acquired, allowing real-time monitoring and quantitative measurements of cell migration.

Figure 3. Live Cell Migration of HT1080 cells. The effect of serum concentrations (0% or 10%) on the migration propensity of HT1080 cells on collagen-coated surface. The results show that the majority of the cells have directed migration towards 10% FCS gradient (Black).

In Vitro Scratch Assay

The scratch assay is a popular method for the study of cell migration⁴. Scaling up this technique has proved challenging, however, making biochemical analysis of the molecular events mediating wound repair difficult. The Cell Comb™ scratch assay addresses the need for a simple tool able to create multiple scratch wounds in a higher-throughput manner.

Product No.	Product Description
17-10191	Cell Comb™ Scratch Assay

Figure 4. Using the CellComb scratch assay, monolayers of NIH3T3 cells were scratched/wounded in a one-direction (left) or two-direction (right) pattern. Over time, migrating cells will fill in the scratched sections and quantified using simple cell counting. Live cell imaging on migrating cells is also possible using the scratch migration assay.

ECM Proteins

Extracellular matrix (ECM) proteins are produced intracellularly and are subsequently secreted into the surrounding cellular medium, actively regulating a diverse range of cell functions including cell adhesion, differentiation, proliferation, migration, invasion and survival. A primary utility of ECMs in in vitro culture is to promote cellular adhesion while maintaining cell viability and maximizing cell proliferation for downstream cell-based applications.

• Collagen
• Laminin
• Fibronectin
• Vitronectin
• Elastin
• Basement Membrane Extract

References

1. Friedl P, Alexander S. 2011. Cancer Invasion and the Microenvironment: Plasticity and Reciprocity. Cell. 147(5):992-1009. https://doi.org/10.1016/j.cell.2011.11.016

2. Lu P, Takai K, Weaver VM, Werb Z. 2011. Extracellular Matrix Degradation and Remodeling in Development and Disease. Cold Spring Harbor Perspectives in Biology. 3(12):a005058-a005058. https://doi.org/10.1101/cshperspect.a005058

3. Chen H. Boyden Chamber Assay.015-022. https://doi.org/10.1385/1-59259-860-9:015

4. Liang C, Park AY, Guan J. 2007. In vitro scratch assay: a convenient and inexpensive method for analysis of cell migration in vitro. Nat Protoc. 2(2):329-333. https://doi.org/10.1038/nprot.2007.30