The Utility and Limitations of Neurosphere Assay, CD133 Immunophenotyping and Side Population Assay in Glioma Stem Cell Research

Wan, Feng; Zhang, Suojun; Xie, Ruifan; Gao, Bao-cheng; Campos, Benito; Herold‐Mende, Christel; Lei, Ting

doi:10.1111/j.1750-3639.2010.00379.x

Cited by 73 publications

(67 citation statements)

References 140 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…Neurosphere cultures are heterogeneous populations of cells; nevertheless, they maintain the phenotype and genotype of the original tumour better than the same cells cultured as adherent cells under serum-containing conditions [15]. Furthermore, the heterogeneity of neurospheres might mirror the in vivo reality [12].…”

Section: Discussionmentioning

confidence: 99%

“…In this purpose, it is necessary to ensure that GSCs have been isolated by at least demonstrating self-renewal over an extended period of time [12,58]. The three studies selected in this paper have used the neurosphere assay as method of GSC isolation; all GSC cultures studied in these three studies were at passage >6 (for a total of 20 cultures), except for eight cultures in the study of Lee et al [15] which were at passage 3 or 5.…”

Section: Discussionmentioning

confidence: 99%

“…A standard in vitro method called "neurosphere assay" has been proposed to enrich cancer stem cells, also called tumour-initiating cells (TICs), from brain tumours [12]. In fact, the neurosphere assay was one of the original methods used to identify putative cancer stem cells within brain tumours [3].…”

Section: Introductionmentioning

confidence: 99%

“…GBM-derived neurosphere cultures are heterogeneous cell clusters that consist of stem cells, various progenitor cells and more differentiated cells. Despite the fact that these assays are currently used as the standard in vitro method for identifying the presence of GSCs, there are some limitations and the reliability is still under debate [12,13]. Indeed, the population of cells isolated by this method represents a heterogeneous rather than a uniform population of cells that are in fact moderately enriched for GSCs [12].…”

Section: Introductionmentioning

confidence: 99%

“…Despite the fact that these assays are currently used as the standard in vitro method for identifying the presence of GSCs, there are some limitations and the reliability is still under debate [12,13]. Indeed, the population of cells isolated by this method represents a heterogeneous rather than a uniform population of cells that are in fact moderately enriched for GSCs [12]. Nonetheless, neurosphere cultures remain informative and important surrogates especially to show a self-renewal phenotype for GSCs versus non-GSCs.…”

Section: Introductionmentioning

confidence: 99%

See 4 more Smart Citations

Identification of OLIG2 as the most specific glioblastoma stem cell marker starting from comparative analysis of data from similar DNA chip microarray platforms

et al. 2014

View full text Add to dashboard Cite

Despite advances in surgical and adjuvant treatments, overall survival of glioblastoma (GBM) patients remains poor. The cancer stem cell concept suggests that a rare stem cell population, called glioma stem cells (GSCs), has high ability to self-renewal leading to recurrence in GBM. The identification of specific markers of GSCs would provide a powerful tool to detect and to characterise them in order to develop targeted therapies. We carried out a comparative analysis based on the identification of inter-study concordances to identify the genes that exhibit at best differential levels of expression between GSC-enriched cell cultures and differentiated tumour cell cultures from independent studies using DNA chip microarray technologies. We finally studied the protein expression of the marker we considered the most specific by immunohistochemistry and semi-quantitative analysis on a retrospective series of 18 GBMs. Of the selected studies, 32 genes were retained. Among them, eight genes were identified to be overexpressed in GSC-enriched cultures compared to differentiated tumour cell cultures. Finally, among the eight genes, oligodendrocyte lineage transcription factor 2 (OLIG2) was characterised by the most different expression level in the "GSC model" compared to the "differentiated tumour cells model". Our approach suggests that OLIG2 is the most specific GSC marker; additional investigations with careful considerations about methodology and strategies of validation are, however, mandatory.

show abstract

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

Identification of OLIG2 as the most specific glioblastoma stem cell marker starting from comparative analysis of data from similar DNA chip microarray platforms

et al. 2014

View full text Add to dashboard Cite

show abstract

The Wnt secretion protein Evi/Gpr177 promotes glioma tumourigenesis

et al. 2011

View full text Add to dashboard Cite

Malignant astrocytomas are highly aggressive brain tumours with poor prognosis. While a number of structural genomic changes and dysregulation of signalling pathways in gliomas have been described, the identification of biomarkers and druggable targets remains an important task for novel diagnostic and therapeutic approaches. Here, we show that the Wnt-specific secretory protein Evi (also known as GPR177/Wntless/Sprinter) is overexpressed in astrocytic gliomas. Evi/Wls is a core Wnt signalling component and a specific regulator of pan-Wnt protein secretion, affecting both canonical and non-canonical signalling. We demonstrate that its depletion in glioma and glioma-derived stem-like cells led to decreased cell proliferation and apoptosis. Furthermore, Evi/Wls silencing in glioma cells reduced cell migration and the capacity to form tumours in vivo. We further show that Evi/Wls overexpression is sufficient to promote downstream Wnt signalling. Taken together, our study identifies Evi/Wls as an essential regulator of glioma tumourigenesis, identifying a pathway-specific protein trafficking factor as an oncogene and offering novel therapeutic options to interfere with the aberrant regulation of growth factors at the site of production.

show abstract