Cell apoptosis of hAMSCs under SC and SF conditions at the indicated time points was accomplished by utilizing the Annexin V and 7-AAD based FCM assay as we previously described [5 (link)]. In detail, hAMSCs under SC and SF conditions were collected and dyed with the commercial FITC Annexin V Apoptosis Detection Kit with 7-AAD (BioLegend, USA) according to the manufacturers’ instructions and detected by FCM assay (BD Biosciences, USA).
As to cell cycle analysis, hAMSCs under SC and SF conditions (preconditioned in SF medium for 48 h) were fixed with 70% cold ethanol (Tianjin, China) after washing with cold 1 × PBS (Gibco, USA). Then, the cells were incubated with the commercial PI/RNase staining solution (BD Pharmingen, USA) at 4 °C for 30 min and detected by using the FACS Canto II (BD Biosciences, USA).
As to cell cycle analysis, hAMSCs under SC and SF conditions (preconditioned in SF medium for 48 h) were fixed with 70% cold ethanol (Tianjin, China) after washing with cold 1 × PBS (Gibco, USA). Then, the cells were incubated with the commercial PI/RNase staining solution (BD Pharmingen, USA) at 4 °C for 30 min and detected by using the FACS Canto II (BD Biosciences, USA).
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