Scratch-Wound Healing Assay for Müller Cell Migration

Cell migration was evaluated through a scratch-wound healing assay (Vera et al., 2021 (link)), with some modifications. In brief, Müller cells were seeded in 24-well plates until they reached a 90% confluence and serum starved for 12 h to prevent cell proliferation. A scratch “wound” was made on the cell monolayer with a 200 µL sterile-pipette tip. “Wound healing” capacity was evaluated at 0, 12, 24, and 48 h after the addition of culture media containing NMDA, control EVs, NMDA EVs, or EGF (100 ng/mL). EGF was used as a positive control since it can induce migration in this cell type (Pena et al., 2018 (link)). The images from these assays were acquired utilizing an inverted optical microscope (Leica, Wetzlar, Germany) with a 10× objective, using an EC3 camera coupled to the microscope and the Leica Application Suite (LAS) software version 3.2.0. Finally, acquired images were analyzed with the free ImageJ (RRID:SCR_003070) software (NIH).

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Carapia A.K., Martinez-Colin E.J., Segura-Villalobos D., Victoria-Chavez R.Y., Lezama I., Martinez-Martinez E, & Lamas M. (2023). Müller Glia to Müller Glia Extracellular Vesicle-Dependent Signaling Induces Multipotency Genes Nestin and lin28 Expression in Response to N-methyl-D-aspartate (NMDA) Exposure. ASN NEURO, 15, 17590914231183272.

Publication 2023

inverted optical microscope

Assays Cell Cell migration Cell proliferation Culture media Microscope Müller cells Nmda Optical microscope Serum Sterile Wound

Corresponding Organization : Center for Research and Advanced Studies of the National Polytechnic Institute

Other organizations : National Institute of Genomic Medicine

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