Flow cytometry analysis and sorting

FcR blocking reagent (Miltenyi Biotec GmbH, Bergisch Gladbach, Germany) was added to 5×10⁵ cells suspended in PBS and incubated at 4°C for 10 min. Subsequently, cells were separately stained with phycoerythrin-conjugated anti-human CD133 or anti-human IgG isotype antibodies (R&D Systems, Inc., Minneapolis, MN, USA) for 30–40 min in 4°C. IgG isotype was used as negative control. Ice-cold PBS was used as washing reagent. Flow cytometry analysis was performed on Accuri™ C6 (BD Biosciences, San Jose, CA, USA) using CFlow software (FCS3.0; BD Biosciences). Flow cytometry sorting was conducted using a BD FACSAria™ I (BD Biosciences).