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Among markers of glioblastoma initiating cells, AC133 has been shown tobe associated with glioblastoma resistance and malignancy. Recently, it was demonstratedthat increasing oxygen tension (pO2) down-regulated AC133 expression in glioblastomacells in vitro. In order to better understand extrinsic factor regulation of AC133,this work aimed to investigate the relationship between cell culture pO2, AC133expression, and tumor development and phenotype. Using treatments with CoCl2 andHIF-1α shRNA knockdowns on non-sorted human primary glioblastoma cells culturedat low (3%) versus high (21%) oxygen tension, we established a responsibilityfor low pO2 in the maintenance of high levels of AC133 expression, with a majorbut non-exclusive role for HIF-1α. We also demonstrated that human glioblastomacells previously cultured under high oxygen tension can lose part of their aggressivenesswhen orthotopically engrafted in SCID mice or lead to tumors with distinct phenotypesand no re-expression of AC133. These observations showed that the specific pO2microenvironment irreversibly impacts glioblastoma cell phenotypes, highlightingthe pertinence of culture conditions when extrapolating data from xenogenic modelsto human cells in their source environment. They also raised AC133 as a markerof non-exposure to oxygenated areas rather than a marker of aggressiveness orlow pO2 niches.

international journal of oncology

In vitro expansion of human glioblastoma cells at non-physiological oxygen tension irreversibly alters subsequent in vivo aggressiveness and AC133 expression