Isolation of Human Glioblastoma Multiforme Primary Cells and analysis of CD133+

Abstract

ObjectiveGlioblastoma multiforme (GBM) is an incurable and the most common type of lethal brain tumors. A transmembrane glycoprotein CD133 has been used as a marker in cancer stem cell analysis.In this study, we aimed isolation of GBM Primary Cells from human GBM tumor tissue which is surgically resected and to determine amount of CD133+ in the primary cultures with using AC133 monoclonal antibody.Material Method : Primer Cell cultureTumor tissues taken from three patients who had GBM diagnosis were separated into 1 mm3 fractions with bistoury within fresh medium. 1ml of DMEM mixture were seeded into T‐25 cm2 culture flasks and incubated at 37°C with 5% CO2.CD133+ stainingCD133+ staining was used as a marker of GBM stem cell marker. Cells were method according to cell surface staining.ResultsWe demonstrated the GBM primary culture isolation of tumor tissues taken from four patients who had GBM diagnosis. We studied three GBM primary cultures such as P1, P2, and P3. We performed amount of CD133+ cells in GBM primary cell cultures by using the method of FCM analysis. According to the FCM analysis results in %1,5 ‐% 1,6 ‐%1,3 are founded respectively.ConclusionFinally, Isolated GBM primary cells provide an invaluable mediate to study how tumors form, progress, and relapse and also to elucidate the underlying deriving cellular mechanisms that eventually could provide understandings to therapeutic options. GBM Stem Cells (GSC) are responsible for tumor generation and treatment resistance in GBM. Identifying GSC cells is an important step in GBM therapy.