It has been postulated that @entity1313 stem cells (TSCs) may be the population of cells responsible for the maintenance and recurrence of @entity463 ( @entity463 ). The purpose of this study was to optimize a reproducible protocol for generating TSCs for their subsequent transfection or transduction. @entity1 GBMs were enzymatically and mechanically dissociated and @entity5 spheres were resuspended in appropriate media and analyzed to ensure they met stem cell criteria. These cells were then transfected with a plasmid or transduced with a viral vector to introduce a previously absent gene and then allowed to form @entity5 spheres. @entity5 spheres were generated from @entity1 GBMs without contamination. These cells met stringent criteria as stem cells, including multipotentiality and self-renewal. High efficiency transfection and transduction of @entity5 spheres was possible, even at the core of the sphere. This allowed for the introduction of new genes to the TSCs, as evidenced by fluorescent microscopy and Western blot analysis. This study is a guide to optimize the generation of @entity1 derived @entity463 spheres without RBC and dead cell contamination. @entity463 TSCs within @entity5 spheres can easily be transfected with plasmids or transduced with a virus. This is important from a therapeutic perspective if gene replacement is to be successful in replacing genes lost in @entity463 progression or to knock down or silence genes that are over-expressed in malignant @entity1313 .

Optimization of XXXX stem cell isolation, transfection, and transduction.

@entity1 :: ('9606', 'Species') :: ['patient', 'Patient'] @entity463 :: ('MESH:D005909', 'Disease') :: ['glioblastoma multiforme', 'GBM', 'GBM tumor'] @entity5 :: ('MESH:D009369', 'Disease') :: ['tumor', 'Tumor'] @entity1313 :: ('MESH:D001932', 'Disease') :: ['brain tumor', 'brain tumors']
@entity463 :: (MESH:D005909,Disease) :: ['glioblastoma multiforme']
