Pi annexinv apoptosis detection kit 1

Cell apoptosis was determined by staining cells with a PI-AnnexinV Apoptosis Detection kit I (BD Biosciences, USA). A total of 1×10⁶ cells were collected and then washed twice with PBS. After the cells were centrifuged at 1500 rpm for 5 min, the cell pellets were resuspended in 1× binding buffer, and the cells were centrifuged again, as previously described, and the stained cells were incubated with 100 μl of 1× binding buffer containing 5 μl AnnexinV/FITC and 10 μL PI for 15 min at room temperature in the dark. Before the flow cytometry analysis, 400 μL of 1× binding buffer was added to each sample.

To measure cell apoptosis, PI-Annexin V Apoptosis Detection Kit I (BD Biosciences, United States) was used according to the manufacturer’s instructions. Briefly, MSCs were collected and washed three times. Then, cell pellets were resuspended in 1 × binding buffer (100 μl) and stained with Annexin V/FITC (5 μl) and PI (10 μl) for 15 min at room temperature in the dark. Afterward, apoptotic events were detected using a flow cytometer (FACS Calibur, BD Biosciences, United States).