Cell migration and invasion play an important role in various physiological and pathological processes like tissue repair, angiogenesis, cancer metastasis, and inflammation. Improper regulation of these cellular processes can lead to the development and progression of several diseases. The cell migration and invasion assays help scientists to understand and study these processes at the cellular and molecular level. These assays allow quantitative measurement of cellular motility and invasion ability under different conditions.
Scratch Cell Migration And Cell Invasion Assay
One of the simplest Cell Migration And Cell Invasion Assay assays used is the scratch wound healing assay. In this assay, a gap or scratch is created in a monolayer of adherent cells using a pipette tip or sterile tip. This creates an artificial wound in the monolayer. Cells are allowed to migrate into the scratch and the migration rate is measured by measuring the narrowing of the scratch gap over time. Images are captured at regular intervals like 0, 12, 24 hours to monitor and quantify the wound closure rate. Parameters like percentage of wound closure and migration rate can be calculated from this assay. It provides a cost-effective way to study the influence of various factors like chemokines, drugs, extracellular matrix on cell migration. However, it does not exactly mimic in vivo conditions.
Boyden Chamber/Transwell Migration Assay
Boyden chamber or transwell migration assay is another widely used assay to measure cell migration. In this assay, cells are seeded on top of an extracellular matrix-coated membrane insert placed within a boyden chamber. The lower chamber contains a chemoattractant like growth factors or serum to induce directed migration of cells through the pores of the membrane. After incubation, cells that have migrated and attached to the bottom side of the membrane insert are fixed, stained and counted under a microscope. The number of migrated cells provides a quantitative measure of cell migration. Transwell assay replicates the process of directed cell migration more closely than scratch wound assay. Various parameters like membrane pore size, extracellular matrix coating can be altered as per experimental requirement.
Spheroid Invasion Assay
The spheroid invasion assay is specifically used to study cell invasion properties. In this assay, multicellular spheroids are formed by culturing cells in non-adherent conditions like hanging drop method or ultra-low attachment plates. These preformed spheroids are embedded in extracellular matrix proteins like matrigel and collagen. Cells from the spheroid edges invade out into the surrounding matrix over time. The degree and extent of radial invasion of cells from spheroids into the matrix correspond to the invasive potential of those cells. This assay mimics the key steps of cell invasion seen during tumor metastasis in vivo. Parameters like invasion area, invasion distance, number of invading cells provide quantitative measure of cell invasion ability under different conditions.
Live Cell Imaging Of Cell Migration And Invasion
Live cell imaging techniques using time-lapse microscopy allow non-invasive and real-time visualization of dynamic cellular processes like migration and invasion over extended periods. In live cell imaging studies of cell migration, cells expressing fluorescent proteins are subjected to scratch wound assay and monitored continuously by capturing images at regular intervals as they migrate into the gap. Similarly, cells within preformed spheroids can be time-lapse imaged as they invade into surrounding extracellular matrix proteins. Live cell imaging provides quantitative data on parameters like migration/invasion speed, persistence of migration, turning angle and morphological changes in migrating cells over time with high spatial and temporal resolution. This helps gain deeper mechanistic insights into regulation of these processes.
Cell migration and invasion assays have contributed significantly to our understanding of various physiological and pathological cellular events at molecular level. Scratch wound assay, Boyden chamber assay, spheroid invasion assay are some commonly employed assays to quantitatively assess migration and invasion ability of cells. Advancements like live cell imaging have enhanced real-time visualization of these dynamic processes. Ongoing development of newer assays will continue to augment research on mechanisms governing movement of cells and their roles in normal tissue homeostasis as well as disease progression.
Get more insights on this topic: https://www.dailyprbulletin.com/cell-migration-and-cell-invasion-assay-new-advancements-in-cell-migration-and-invasion-assays-enhancing-understanding-of-cell-behavior-2/
Author Bio
Vaagisha brings over three years of expertise as a content editor in the market research domain. Originally a creative writer, she discovered her passion for editing, combining her flair for writing with a meticulous eye for detail. Her ability to craft and refine compelling content makes her an invaluable asset in delivering polished and engaging write-ups. (LinkedIn: https://www.linkedin.com/in/vaagisha-singh-8080b91)
*Note:
1. Source: Coherent Market Insights, Public sources, Desk research
2. We have leveraged AI tools to mine information and compile it