Cd73 percp cy5

Mice were intracranial implanted with GL261 WT or GL261-CD73 cells as described above. When mice showed signs of neurological deficit, they were transcardially perfused. The tumor-bearing hemispheres were minced into small pieces with scalpels, washed, and passed through a 70 μm cell strainer. The single-cell suspension was stained first with a fixable viability dye (Thermo Fisher Scientific), before staining with antibodies specific for murine CD45-BV510 (RRID:AB_2563061) and CD73-PerCP-Cy5.5 (RRID:AB_11219403) (both BioLegend), and measured using FACS Celesta (BD Biosciences). The data analysis was performed with FlowJo software.

The immune phenotype of CIMVs-MSCs was analyzed by immunostaining with monoclonal antibodies: CD90-PE-Cy5 (328112; BioLegend, San Diego, CA, USA), CD29-APC (2115040; Sony, San Jose, CA, USA), CD44-APC/Cy7 (103028; BioLegend, San Diego, CA, USA), CD73-PerCP/Cy5.5 (344014; BioLegend, San Diego, CA, USA). CIMVs were analyzed by flow cytometry (BD FACS Aria III, BD Bioscience, San Jose, CA, USA), and the 405 nm laser was used for better a resolution of CIMVs-MSC.