Unravel molecular profil of astrocyte-like and oligodendrocyte-like cells in low grade glioma cell line LGG275

Astrocytoma are known to presents a cellular heterogeneity consisting of three identified subpopulation: "astrocyte-like", "oligodendrocyte-like" and "NPC-like" cells. Properties of these subpopulation remains unclears, specially due to the fact that no protocol allows isolation of each subpopulation. In these study, we develop a protocol to efficiently sort two populations, "astrocyte-like" and "oligodendrocyte-like" cells, by using 2 astrocyte markers; CD44 and GLAST. Overall design: After 5 days culture without growth factors, cells were stained with CD44 and GLAST antibodies, then sorted to isolate astrocyte-like CD44+GLAST+ (POS) cells, and oligodendrocyte-like CD44-GLAST- (NEG) cells. RNA extraction of POS, NEG or unsorted (U) cells were performed directly after cell sorting