Cck 8 reagent

To evaluate cell proliferation on BiOCl nanosheets, bone marrow mesenchymal stem cells (BMSCs, CRL-12424, ATCC, USA) were cultured with BiOCl and analyzed using the Cell Counting Kit-8 (CCK-8). Briefly, BMSCs were seeded in 96-well cell culture plates at densities of 2500 ​cell per well in DMEM containing 10% FBS at 37 ​°C in a humid atmosphere of 5% CO₂. After 24 ​h incubation, the cells were incubated with BiOCl nanosheets of different concentrations. Cells, cultured in the medium without nanomaterials, were used as the negative control. After an additional 24 ​h of incubation, 10 ​μL of the CCK-8 reagent (Dojindo, Japan) was added, and the culture was incubated another 2 ​h at 37 ​°C. The optical density was measured using a spectral scanning multimode reader (Thermo Scientific Varioskan Flash, USA) at a 450 ​nm wavelength. For light illuminated groups, cells were cultured with nanosheets in the same manner as the aforementioned process. After cells were incubated with BiOCl nanosheets for 24 ​h, the nanosheets and blank controls were illuminated at 365 ​nm with a LED source at a power density of 4.25 ​mW ​cm⁻² for 30 ​min, then the same condition with normal CCK-8 tests.

Cell viability was assessed by a cell counting kit-8 (CCK-8). Briefly, PKH26-labelled MenSCs (PKH26-MenSCs) and unlabelled MenSCs were separately seeded at a density of 1000 cells per well in 96-well plates. Ten microliters of CCK-8 reagent (Dojindo Laboratories, Japan) was added to 6 wells of each group every 24 h, and the cells were incubated for 1 h at 37 °C in 5% CO_2. The absorbance at 450 nm was measured by a spectrophotometer (Beckman Coulter, United States), and the growth curve was drawn to compare cell proliferation ability between the two groups. To induce osteogenic differentiation, PKH26-MenSCs and unlabeled MenSCs were cultured in commercially available osteogenesis differentiation medium (Cyagen Biosciences, United States). On day 21, cells were stained with alizarin red S. To induce adipogenic differentiation, each group of cells was cultured in a commercially available adipogenesis differentiation medium purchased from Cyagen. On day 21, lipid droplets were visualized by oil red O.

Cell Counting Kit-8 (CCK-8), colony formation and EdU assays were performed to detect the proliferation of glioma cells. CCK-8 reagent (10μl/well, Dojindo Laboratories, Dojindo, Japan) was added to the plate and cells were cultured for 1.5 hrs. Then, the absorbance of the samples at 450 nm was detected by microplate reader (Bio-Rad, Hercules, CA, USA). For colony formation assay, glioma cells were seeded in six-well plates (200 cells per well) and incubated for 2 weeks until the visible colonies were formed. The colonies with crystal violet staining were counted. The EdU assay was carried out using the Cell-Light™ EdU Apollo^®488 In Vitro Imaging Kit (RioBio) following the manufacturer’s recommendations.