hello. I am a student studying MTT assay and Wound healing.
Studying MTT and Wound healing, I learned that MTT measures cell viability and Wound healing measures cell proliferation rate and inhibition of migration.
However, I thought that it could be said that cell migration was inhibited by the inhibition of cell viability.
Can you tell me the exact difference between MTT and Wound healing?
Or do you have a paper to refer to when studying the difference between MTT and Wound healing?
thank you.
Dear Eunji Han,
Lets first discuss characteristic features we can measure and then we can move on to methods. You may measure cell viability (total number of living cells), proliferation rate (increase in the number of cells over time), cell death etc. Then you have methods: you may utilize an MTT, XTT, SRB assays or just count the number of living cells to assess viability. You may use nucleotide incorporation assays or observe the increase in number of cells over time to measure proliferation rate. And there are various specific assays to measure similar characteristic features. An MTT assay is just an approach to measure cell viability. It measures cellular metabolism as an indicator of cell viability. Other approaches include measuring total protein/DNA or counting dyed/undyed cells.
Depending on your experimental design, proliferation rate may mean wound healing. A classical wound-healing assay relies on observation cell proliferation over time (https://en.wikipedia.org/wiki/Wound_healing_assay). The same method may be utilized as a migration assay as well.
I recommend you to read our paper on topic:
Article From viability to cell death: Claims with insufficient evide...
MTT assay measures the activity of the mitochondria converting MTT to formazan. The wound healing assay is used to see if a given conditions increases or decreases cell migration. Cells can be viable and have reduced migration. Migration and metabolic activity are two distinct parameters.
Hello Eunji Han
The MTT assay is used to measure cellular metabolic activity as an indicator of cell viability, proliferation, and cytotoxicity. This colorimetric assay is based on the reduction of MTT to purple formazan crystals by metabolically active cells. The viable cells contain NAD(P)H-dependent oxidoreductase enzymes which reduces the MTT to formazan. The insoluble formazan crystals are dissolved using a solubilization solution and the resulting-colored solution is quantified spectrophotometrically. The darker the solution, the greater the number of viable, metabolically active cells.
On the other hand, Wound Healing assay is a standard in vitro technique for probing collective cell migration. In this assay, a cell-free area is created in a confluent monolayer by physical exclusion or by removing the cells from the area through mechanical, thermal, or chemical damage. The exposure to the cell-free area induces the cells to migrate into the gap. In this assay it is essential to inhibit proliferation of cells because closure of the wound should happen only due to cell migration and not by cell proliferation. For this reason, the cells need to be starved for at least 16 hours before the scratch (wound) is created. You may provide the cells with 0.5% serum containing media which may help prevent cell death but at the same time inhibit cell proliferation.
So, MTT assay measures cell viability and is mostly used to check the cytotoxic effects of compounds under study. Wound healing assay measures the basic cell migration parameters such as speed, persistence, and polarity. Cells at the edge of the created wound polarize and migrate into the wound space. This assay does not require the use of specific chemo attractants or gradient chambers, and it generates a strong directional migratory response. It is most reliably analyzed when performed using time-lapse imaging, which can also yield valuable cell morphology/protein localization information.
In short, MTT assay checks cell viability and Wound Healing assay checks cell migration in terms of speed, persistence and polarity.
Best.
A wound healing assay is a laboratory technique used to study cell migration and cell–cell interaction. This is also called a scratch assay because it is done by making a scratch on a cell monolayer and capturing images at regular intervals by time lapse microscope. But the MTT Reagent yields low background absorbance values in the absence of cells. For each cell type the linear relation- ship between cell number and signal produced is established, thus allowing an accurate quantification of changes in the rate of cell proliferation
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