by Cell migration is a fundamental process that occurs both during normal physiological conditions such as embryonic development, tissue repair, and immune response as well as in various pathologies including cancer metastasis, atherosclerosis and rheumatoid arthritis. The ability of cells to migrate and invade surrounding tissues is crucial for numerous biological mechanisms. Improper regulation of cell migration can enable conditions such as cancer to occur or exacerbate inflammatory diseases. Therefore, being able to study and analyze cell motility mechanisms provides great insight into normal function as well as disease progression.
Methods For Measuring Cell Migration And Cell Invasion Assay
Several methods are commonly utilized for evaluating and quantifying Cell Migration And Cell Invasion Assay ability in vitro. The basic premise involves establishing a defined population of cells in one area and then measuring how far and how many cells are able to migrate over a set period of time into an open region or towards chemoattractants and chemorepellents. Some of the assays that are widely employed include:
Wound Healing or Scratch Assay: This most simplistic technique involves creating an artificial wound or gap in a cell monolayer using a pipette tip or similar instrument. Microscopic images are captured at initial time point and at various intervals as cells migrate to close the wound gap over hours or days. The changed wound area is calculated which directly correlates to migratory ability.
Boyden or Transwell Chamber Assay: Cells are seeded on microporous membrane inserts that are placed within wells containing chemoattractants or control media. Cells that migrate towards the lower side of membrane are fixed, stained and counted under microscope or with colorimetry. A higher number of cells passing through pores indicates increased directed cell migration.
Live-Cell Imaging: Advance live-cell imaging platforms allow continuous monitoring of fluorescently labeled cells in real-time over many hours as they crawl and make protrusions. Parameters like migration speed and directionality can be tracked using specialized analysis software. This provides a dynamic view of migration in response to extracellular factors.
Scratch Wound Assay Variants: Modifications using microfluidic devices or reusable silicone membranes permit application of gradients and evaluation of collective cell migration in response to precisely regulated environmental cues mimicking in vivo conditions.
Evaluation Of Cell Invasion
While cell migration refers to movement on surfaces, cell invasion involves migration through extracellular matrices and basement membranes – a characteristic of metastatic cancer cells and other cell types. Dedicated invasion assays are developed to assess this key attribute of cells.
Commonly Used Methods Include:
Matrigel Invasion Chamber Assay: Matrigel which mimics the three-dimensional basement membrane structure is coated on the membrane inserts. Cells seeded on top are allowed to invade through pores towards a chemoattractant. Invasive cell number counted after removal of non-invasive cells.
Spheroid Invasion Assay: Spheroids of tumor or other cells are embedded in 3D matrices like Matrigel or collagen gels. Time-lapse imaging allows monitoring their invasive protrusions and penetration into surrounding matrix over time. Computational analysis provides invasion metrics.
Collagen Contraction Assay: Cells are embedded in floating collagen gel matrices. Invasion ability is determined by collagen gel shrinkage due to cellular contraction force. Increased invasion causes greater gel area reduction.
Microfluidic Invasion Assays: Microengineered biomimetic invasion models containing gradients and interstitial flow conditions provide a more native physiological microenvironment to study directed three-dimensional multicellular invasion of tumor spheroids or strands. Real-time microscopic observations and quantitation of invasion parameters are possible.
These dedicated invasion assays along with migration assays have significantly enhanced understanding of mechanisms governing cell motility and factors determining their metastatic potential or inflammatory behavior under normal as well as pathological conditions. They serve as invaluable tools in basic cell biology research as well as drug discovery efforts targeting motility-dependent diseases.
Various cell culture methods widely applied to study and quantify two important modes of cell motility – migration and invasion – were described. Cell migration assays including wound healing, transwell chambers and live-cell imaging techniques measure horizontal movement of cells across surfaces. Matrigel and collagen invasion models embedded in three dimensions assess vertical invasion through extracellular matrices, a hallmark of metastatic ability. Combined use of these assays continues providing novel insights into molecular and biophysical determinants of cell motility central to development, immunity, wound repair as well as pathology of cancer and other diseases.
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1. Source: Coherent Market Insights, Public sources, Desk research
2. We have leveraged AI tools to mine information and compile it
About Author - Alice Mutum
Alice Mutum is a seasoned senior content editor at Coherent Market Insights, leveraging extensive expertise gained from her previous role as a content writer. With seven years in content development, Alice masterfully employs SEO best practices and cutting-edge digital marketing strategies to craft high-ranking, impactful content. As an editor, she meticulously ensures flawless grammar and punctuation, precise data accuracy, and perfect alignment with audience needs in every research report. Alice's dedication to excellence and her strategic approach to content make her an invaluable asset in the world of market insights. LinkedIn
