Explore the molecular profile and properties of quiescent (P27+) and mitotic (P27-) glioma cell in diffuse low grade astrocytoma cell lines LGG275

Astrocytomas are known to present a cellular heterogeneity consisting of three identified subpopulations: "astrocyte-like", "oligodendrocyte-like" and "NPC-like" cells. It has also been shown that they contain a subpopulations of quiescent, or neural G0 cells, representing more than 90% of the tumor cells. The properties of this cell remain unclear due to the fact that it is difficult to isolate them. We used mVenus/P27 vector expression, constructed by Oki et al (2014), where P27 is fused to mVenus to highlight quiescent cells in LGG275 cell lines, a slow-growing astrocytoma. Then we used this reporter system to sort the mVenus/P27 expressing cells (quiescent) and the negative cells (mitotic) in aims to explore the properties of these two populations and validate the mVenus/P27 tool. Overall design: LGG275 cells were infected with a lentivirus containing mVenus/P27 construction, where mVenus is fused to P27. Then, cells were sorted based on their expression of mVenus. mVenus pos cell were P27 pos (quiescent) cells. mVenus neg cells were P27 neg (mitotics) cells.