Wound Healing Assay with Extracellular Vesicles

A375, 501Mel cells and NHDFs were seeded at a density of 3 × 10⁴, 3.5 × 10⁴ and 16.7 × 10⁴ cells/well, respectively, in collagen I-coated (200 µg/mL, Merck Millipore, Burlington, MA, USA) 96-well plates (Essen Imagelock, Essen Bioscience, Ann Arbor, MI, USA). Then, 24 h after seeding, when cells had reached 80–90% confluence, a wound was scratched across each well with the Cellplayer 96-well woundmaker (Essen Bioscience, Ann Arbor, MI, USA). After the scratching, 5 µg nEVs or hEVs were added per well. Migration and invasion were studied as previously described in [78 (link)].

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Walbrecq G., Lecha O., Gaigneaux A., Fougeras M.R., Philippidou D., Margue C., Tetsi Nomigni M., Bernardin F., Dittmar G., Behrmann I, & Kreis S. (2020). Hypoxia-Induced Adaptations of miRNomes and Proteomes in Melanoma Cells and Their Secreted Extracellular Vesicles. Cancers, 12(3), 692.

Publication 2020

Cellplayer 96-well woundmaker

Cells Collagen i Hevs Wound

Corresponding Organization : University of Luxembourg

Other organizations : Luxembourg Institute of Health

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Variable analysis

independent variables

Type of extracellular vesicles (nEVs or hEVs)

dependent variables

Cell migration
Cell invasion

control variables

Cell density for A375 cells (3 × 10^4 cells/well)
Cell density for 501Mel cells (3.5 × 10^4 cells/well)
Cell density for NHDFs (16.7 × 10^4 cells/well)
Collagen I coating (200 µg/mL)
96-well plates (Essen Imagelock)
Time after seeding (24 h)
Confluence level (80-90%)
Wound scratch method (Cellplayer 96-well woundmaker)

positive control

Not explicitly mentioned

negative control

Not explicitly mentioned

Annotations

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