The Role of CSCs/CCs Derived from GBM Patient in Switching of the Healthy Macrophage from M1 to M2

Glioblastoma multiform (GBM) is the most aggressive malignant primary brain tumor.Many factors contribute to the inevitable recurrence ofGBM and its resistance to current therapeutics. One of the most commonly detected phenomena in GBM is the abundant macrophage infiltration without apparent phagocytic activity. Accumulating evidences have suggested that the tumor-associated macrophage(TAM) infiltration can be linked to the poor prognosis in GBM. There are controversial reports about the origin, the recruitment, and the functions of the TAMs in GBM. Depending on macrophages/microglia receive which signals from their surroundings, their functionalphenotype would be changed. They could be polarized to M1 phenotype in response to an inflammatory condition and produce pro-inflammatory factors such as IL-6 or IL-12. In contrast, in anti-inflammatory condition they would be polarized to M2 phenotype,which characterized by wound healing, angiogenic and pro-tumorigenic properties. So, in this study, we are investigating the role of cancer stem cells/cancer cells derived from GBM patient in switching of the healthy macrophage from M1 to M2. Materials and Methods:Following surgical resection, human brain tumor biopsy was dissociated by mechanical and chemical digestion. Cell pellets were resuspended and cultured to specefic medium. Finally, to characterize the isolated cells, immunostaining was performed. Also we isolated human CD14+ cells from leukoreduction system chamber and then monocytes differentiate into macrophages was done. Flow cytometric analysis of these cells was performed with specific markers. Results: Isolated CSCs/CC from GBM express Nestin, Sox2, Cd133 and Ki67 markers. CD14+ monocytic cells were isolated by CD14 Microbeads. IFN-g was used for macrophage polarization. Purity of monocytes and macrophage were assessed by flow cytometry. Conclution: A better understanding of the interaction of anticancer therapieswith innate immunity, and TAMs in particular, may have the way to better patient selection and innovative combination of conventional approaches with immunotherapy.