Cell Culture, Migration, and Proliferation Assays

The established cell cultures were cultivated until the confluency grade became 80%; then, the cells were removed from the flask surfaces by using 0.25% trypsin solution (Thermo Fisher, Waltham, MA, USA) and reseeded on new flasks. As a cell culture media was used high-glucose DMEM with 1mM Sodium Pyruvate and 300 mg/L L-glutamine (Thermo Fisher, Waltham, MA, USA), supplied with 10% Fetal Bovine serum (FBS) (Thermo Fisher, Waltham, MA, USA), penicillin and streptomycin (Thermo Fisher, Waltham, MA, USA).
The migration and proliferation assays were considered by using a time-lapse Cell IQ machine, which is described by Narkilahti et al. [62 (link)]. This system acts as a hardware phase-contrast microscope and CO₂ incubator assembly and a software complex designed for image recognition for the quantification of proliferation and migration image data.