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The reduction of cell coupling when confluence is achieved supports the proposed role of GJIC in regulating astrocyte migration.
Once confluence is achieved, trypsin and EDTA are used to disrupt adherence, and MSCs are re-plated within a larger number of flasks [ 30].
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Cells were supplemented with fresh improved medium every 3 4 days, depending on the requirements, until confluence was achieved.
After the cells' confluence was achieved in the microchambers, the novel method of cells' passaging in the designed microdevice was developed and successfully tested.
After confluence was achieved, the TEER value of cells was measured to verify the presence of tight junction.
hMSCs were cultured and maintained in DMEM with 10% fetal calf serum and 1% penicillin, at 37 °C in 5% CO2 humidified atmosphere till 90% confluence was achieved.
RADMSCs and HeLa were cultured on coverslips, and the medium was changed every 2 days, until 85% cell confluence was achieved.
To study the cellular internalization efficiency of the silica-coated CdSe QDs in vitro, RADMSCs and HeLa cells were cultured on coverslips and the medium was changed on every 2 days, till 85% cell confluence was achieved.
These cells were thawed on the same day and cultured in 6-well plate in the presence of hygromycin B until 80% confluence was achieved.
For BrdU labeling, SRA overexpressing or control ST2 preadipocytes were plated on chamber coverglass slides and maintained in the ST2 cell media described above until one day after confluence was achieved.
After one passage (p1), NSCs were seeded at a density of 5×104 in 35 mm plates pre-coated with poly-L-ornithine and fibronectin (Sigma, St . Louis MO, USA) and expanded in N2 media until >60% confluence was achieved.
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