Isolation and Expansion of Human Glioblastoma Multiforme Tumor Cells Using the Neurosphere Assay

• DOI: 10.3791/3633
• Publication Date: 2011-11-07T22:07:28Z
• Authors: Hassan Azari; Sebastien Millette; Saeed Ansari; Maryam Rahman; Loic P. Deleyrolle; Brent A. Reynolds
• Keywords: Neurosphere; Brain tumor

Stem-like cells have been isolated in tumors such as breast, lung, colon, prostate and brain. A critical issue all these tumors, especially glioblastoma mutliforme (GBM), is to identify isolate tumor initiating cell population(s) investigate their role formation, progression, recurrence. Understanding populations will provide clues finding effective therapeutic approaches for tumors. The neurosphere assay (NSA) due its simplicity reproducibility has used the method of choice isolation propagation many this cells. This protocol demonstrates culture expand stem-like surgically resected human GBM tissue. procedures include an initial chemical digestion mechanical dissociation tissue, subsequently plating resulting single suspension NSA culture. After 7-10 days, primary neurospheres 150-200 μm diameter can be observed are ready further passaging expansion.