96 well cell culture cluster plates

To value the capacity of BM-MSC exosomes to facilitate NPC proliferation, the Cell Counting Kit-8 assay was used. NPCs were seeded on 96-well cell culture cluster plates (Corning Inc., Corning, NY, USA) at a concentration of 2 × 10³ cells/well in volumes of 100 μl and grown overnight. The duration of BM-MSC exosome interaction with NPCs was grouped into 3, 6, 9 and 12 days and a control group (NPCs alone without BM-MSC exosomes). To eliminate the detection error, all groups (including the control group) were cultured for 12 days under the same condition and the BM-MSC exosomes (50 μg/ml, resuspended in 100 μl DMEM/F-12 medium) were added to the NPCs at the due time according the timing of the experimental design. Finally, 10 μl of Cell Counting Kit-8 reagents (Dojindo, Kumamoto, Japan) was added to each well at the due time points and then incubated for another 2 hours at 37 °C in the dark. The optical density (OD) value was measured at 450 nm, and three trials were performed to obtain the average values. The relative proliferation rate to control was used to create the cell growth chart and examined using one-way ANOVA.

Cell counting kit-8 (CCK8) assay was performed to evaluate cell proliferation. A549 cells were seeded into 96-well cell culture cluster plates (Corning, USA) at 2×10⁴ cells/well, cultured overnight, and then transfected with DISC1-siRNA2 and negative control siRNA. CCK-8 reagents (Dojindo, Japan) were added to each well at different time points, and wells were incubated for an additional 2 h at 37°C in the dark. Absorbency was measured at 450 nm (reference 650 nm) with a microplate reader (Bio-Rad, USA). Experiments were repeated at least three times.