title: Culture and isolation of brain tumor initiating cells.
abstract: This unit describes protocols for the culture and isolation of brain tumor initiating cells (BTIC). The cancer stem cell (CSC) hypothesis suggests that tumors are maintained exclusively by a rare fraction of cells that have stem cell properties. We applied culture conditions and assays originally used for normal neural stem cells (NSCs) in vitro to a variety of brain tumors. The BTIC were isolated by fluorescence activated cell sorting for the neural precursor cell surface marker CD133. Only the CD133(+) brain tumor fraction contains cells capable of sphere formation and sustained self-renewal in vitro, and tumor initiation in NOD-SCID mouse brains. Therefore, CD133(+) BTICs satisfy the definition of cancer stem cells in that they are able to generate a replica of the patient's tumor and they exhibit self-renewal ability through serial retransplantation. This established that only a rare subset of brain tumor cells with stem cell properties are tumor-initiating, and, in this unit, we describe their culture and isolation.
authors.affiliation: McMaster University, Hamilton, Ontario, Canada.
authors.forename: Monika
authors.initials: M
authors.lastname: Lenkiewicz
authors.forename: Na
authors.initials: N
authors.lastname: Li
authors.forename: Sheila K
authors.initials: SK
authors.lastname: Singh
mesh_terms: D000071916:AC133 Antigen; D000818:Animals; D015703:Antigens, CD / Q000096:biosynthesis; D001921:Brain / Q000378:metabolism; D001932:Brain Neoplasms / Q000378:metabolism / Q000473:pathology*; D018929:Cell Culture Techniques / Q000379:methods*; D002469:Cell Separation / Q000379:methods; D005434:Flow Cytometry / Q000379:methods; D006023:Glycoproteins / Q000096:biosynthesis; D006801:Humans; D008280:Magnetics; D051379:Mice; D016688:Mice, Inbred NOD; D016513:Mice, SCID; D014411:Neoplastic Stem Cells / Q000166:cytology*; D010455:Peptides; D011994:Recombinant Proteins / Q000737:chemistry; D014407:Tumor Cells, Cultured / Q000166:cytology*
chemical_list: D000071916:AC133 Antigen; D015703:Antigens, CD; D006023:Glycoproteins; C000606023:PROM1 protein, human; D010455:Peptides; C000606024:Prom1 protein, mouse; D011994:Recombinant Proteins