Isolation of Glioma-Initiating Cells for Biological Study.
Glioblastoma multiforme (GBM, WHO grade IV astrocytoma) is the most common and lethal primary brain tumor in adults, with an average survival of slightly more than 1 year after initial diagnosis. GBMs display significant heterogeneity within the tumor mass, among which a subpopulation of cells called glioma-initiating cells (GICs) is responsible for tumorigenesis and resistance to conventional therapies. Therefore, understanding the mechanism underlying the biological properties of GICs would help develop better therapies to target this population for GBM treatment. This protocol provides detailed procedures to isolate GICs and non-GICs from patient's specimen and glioma xenografts, which serves as the first step for the biological studies of GICs. Upon separation of GICs and non-GICs, a series of studies, such as expression profiling and functional screen, etc., can be performed to identify signal pathways responsible for the malignant nature of GICs. Besides, translational studies can also be conducted to examine drug responses of GICs. In sum, isolation of GICs with reliable methods will provide the basis for the further biological studies.
Department of Pharmacology and Cancer Biology, Duke University School of Medicine, Duke University Medical Center, C218 LSRC, Box 3813, Durham, NC, 27710, USA.
Jing
J
Hu
Department of Pharmacology and Cancer Biology, Duke University School of Medicine, Duke University Medical Center, C218 LSRC, Box 3813, Durham, NC, 27710, USA.
Geoffrey
G
Markowitz
Department of Pharmacology and Cancer Biology, Duke University School of Medicine, Duke University Medical Center, C218 LSRC, Box 3813, Durham, NC, 27710, USA. xiao.fan.wang@duke.edu.
Xiao-Fan
XF
Wang
D015703:Antigens, CD / Q000378:metabolism; D014408:Biomarkers, Tumor / Q000378:metabolism; D002469:Cell Separation / Q000379:methods*; D005434:Flow Cytometry; D005910:Glioma / Q000473:pathology*; D006801:Humans; D014411:Neoplastic Stem Cells / Q000473:pathology*
D015703:Antigens, CD; D014408:Biomarkers, Tumor
CD133; CD15; Glioblastoma; Glioma-initiating cells