Cell Migration and Invasion

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Cell migration plays a vital role in multiple biological and pathological processes, including inflammation, wound healing, cell differentiation and cancer metastasis. Modulating this directed movement offers an effective approach across multiple therapeutic areas.

Directed Cell Migration - Chemotaxis

Chemotaxis is the directed migration of cells in response to a stimulus along a concentration gradient. Tracking chemotaxis is commonly used to approach questions on inflammation and cancer. Historically, assays assessing chemotaxis have utilized modified Boyden chambers that consist of upper and lower chambers separated by a porous filter. However, these assays are of limited utility, typically requiring a large number of cells and only allowing for an endpoint measurement rather than providing information on the kinetics of cell migration.

At Charnwood Discovery, we utilize our Incucyte SX5 live-cell imager to measure chemotaxis through a porous membrane in a 96-well plate format. This allows the development of image-based, kinetic chemotaxis assays, as well as being able to visually monitor and track cell health using fluorescent viability dyes. This approach gives a detailed view of the migration process and its regulation. It also helps separate the effects of compound treatment on chemotaxis from cytotoxicity artifacts.

Red-labelled HT-1080 cells migrate from serum-free medium to medium containing 10% FBS. Left – in this video cells can be seen migrating through regularly spaced pores, images captured every 5 minutes. Right – as per the left-hand video with the Incucyte SX5 analysis mask overlayed.

Here we present example chemotaxis data from our lab. Using the Incucyte SX5, we monitored the chemotaxis of HT-1080 NucLight Red cells (a fibrosarcoma cell line with a red nuclear protein). These cells were treated with a serial dilution of Cytochalasin D. The graphs below demonstrate clear concentration-dependent inhibition of chemotaxis by Cytochalasin D.

Cell migration_concentration-dependent graph_normalized
Cell migration_Dose response graph

Cytochalasin D inhibits chemotactic migration of HT-1080 cells in a concentration-dependent manner. Left – normalized cell count on the bottom of the membrane over time. Right – inhibition of chemotaxis by cytochalasin D after 48 hours.

Wound Healing

Traditional scratch-wound healing assays, commonly used in oncology to assess metastasis mechanisms, present several challenges. Manual scratching of individual wells and capturing images across multiple time points is labor-intensive. This method introduces variability due to inconsistent scratches and limits throughput.

Using our live-cell imaging instrument, the Incucyte SX5, we continuously monitor cell migration into the scratch wound, capturing the kinetics of cell motility. Trainable algorithms ensure consistent analysis. The Sartorius 96-well Woundmaker Tool enhances throughput and reduces variability by simultaneously creating uniform scratches in each well of a 96-well plate. The video below demonstrates a scratch wound’s recovery over two days and tracks migration inhibition using this technology.

Cell migration_wound density graph

Cells migrate to heal a scratch wound made in a monolayer of red-labelled HT-1080 cells. Left – this video demonstrates how the Incucute tracks wound healing over time, the initial scratch wound is coloured blue, while the current wound is labelled orange. Right – cytochalasin D treatment inhibits cell migration and wound healing.


We can utilize both of the aforementioned assay setups to investigate cell invasion by incorporating an extracellular matrix (ECM). This allows us to study cells’ ability to invade through an ECM in response to changes in cell density or a chemical gradient. We can compare invasive properties across different cell types, culture conditions and ECM compositions by measuring  relative levels of invasion against controls and non-invasive cells under selected experimental conditions. Furthermore, we can evaluate the impact of compounds on invasion versus migration within the same microplate by including or excluding ECM.

Designing Assays for Success

Assay development is a critical factor underpinning the success of any drug discovery program. Every step in the protocol must be meticulously evaluated to ensure reliability and accuracy.

Our focus, whether building from literature or using existing assays, is always on ensuring the assay is fit for purpose.

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