Met-Induced Migration in Scratch Assay

The wound healing (scratch) test was used to assess the effect of Met on cell migration. For this purpose, the cells were cultured in 24-well Corning plates (1 × 10⁵ cells/ well) and an automatic scratch was made in the middle of each well using the AutoScratch ™ Wound Making Tool provided by BioTek^® Instruments Inc., Winooski, VT, USA as recommended by the manufacturer. Cells were then stimulated with the five Met concentrations (0.5, 1, 1.5, 2 and 2.5 mM) for a 24-hour interval. Photographs (4 × magnification) were taken both at the beginning of the experiment (0 h) and after the stimulation was completed (24 h) using Cytation 1 and were processed using Gen5 ™ Microplate Data Collection and Analysis Software (BioTek^® Instruments Inc., Winooski, VT, USA). The effect of Met on cell migration was quantified by calculating the migration rate, expressed as a percentage, using the formula described above [19 (link)].

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Cozma G.V., Apostu A., Macasoi I., Dehelean C.A., Cretu O.M., Dinu S., Gaiță D, & Manea A. (2022). In Vitro and In Ovo Evaluation of the Potential Hepatoprotective Effect of Metformin. Medicina, 58(6), 705.

Publication 2022

AutoScratch ™ Wound Making Tool Gen5 ™ Microplate Data Collection and Analysis Software

Cell migration Cells Wound

Corresponding Organization : Victor Babeș University of Medicine and Pharmacy Timișoara

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Variable analysis

independent variables

Met concentrations (0.5, 1, 1.5, 2 and 2.5 mM)

dependent variables

Cell migration rate (expressed as a percentage)

control variables

Cell culture in 24-well Corning plates
Initial cell density (1 × 10^5 cells/ well)
Automatic scratch made in the middle of each well using the AutoScratch ™ Wound Making Tool
Stimulation duration (24-hour interval)
Photographs taken at 0 h and 24 h using Cytation 1 and processed using Gen5 ™ Microplate Data Collection and Analysis Software

controls

No positive or negative controls were explicitly mentioned in the protocol.

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