Transcriptome Changes in Glioma Cells Cultivated under Conditions of Neurosphere Formation

Glioma is the most common and heterogeneous primary brain tumor. The development of the new relevant preclinical models is necessary. As research moves from cultures of adherent gliomas to a more relevant model, neurospheres, it is essential what changes cells undergo at the transcriptome level. Cultivation of primary cultures of gliomas in a medium without serum, but with the addition of growth factors - bFGF, EGF and nutritional supplements, allows the formation of neurospheres enriched in cells with characteristic features of tumor stem cells: the potential for unlimited self-renewal and the ability for readily differentiate to into neurons and glial cells.For comparative analysis of the transcriptome, RNA-seq of 3 human glioma patients-derived cell cultures (BR1, MG1 and BR2) and 2 immortalized human glioma cell lines (U343 MG and U87 MG) in the adhesive state and in the state of neurospheres was performed. We obtained glioma patients-derived cell cultures from 3 solid primary brain tumors (2 glioblastomas and 1 diffuse astrocytoma: BR1, MG1, and BR2, respectively). We cultivated glioma cells in adherent state and in conditions that favored the formation of neurospheres and obtained 5 pairs of adherent and neurosphere cultures. From the obtained human glioma cell cultures, we performed RNA isolation, construction of cDNA libraries, and massive parallel sequencing on the NextSeq Illumina 1500 platform.