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. 2022 Apr 23;23(9):4670. doi: 10.3390/ijms23094670

Figure 4.

Figure 4

Overview of A2B5 expression in glioma highlighting A2B5 as a GBM CSC marker. (A) A2B5+ bipolar OPC/O-2A cells are observed in all glioma types but harbor distinct migration and proliferation capacities, depending on the WHO grade of glioma (pilocytic astrocytoma—PA, grade 1, versus glioblastoma—GBM, grade 4). In primary explant culture assays, A2B5+ cells from PA did not proliferate (few A2B5+/BrdU+ cells); they had a bipolar appearance with very long cell processes, one of them being in close contact with the explant. In contrast, A2B5+ cells from GBM migrated far away from the explant and were dividing (numerous A2B5+/BrdU+ cells). (B) We and others have generated A2B5+ cancer stem cell GBM cell lines; here we summarize our procedure, from (1) tumor resection and mechanic and enzymatic dissociation, (2) A2B5 immunomagnetic cell sorting (A2B5+ cell fraction was about 20% of the single cell suspension), (3) culture in a serum free stem-cell permissive medium to form gliomaspheres and (4) demonstration that only A2B5+ cell fraction gives rise to tumor development after stereotactic xenograft in mice.