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Molecular and phenotypic characterisation of paediatric glioma cell lines as models for preclinical drug development.

Bax DA, Little SE, Gaspar N, Perryman L, Marshall L, Viana-Pereira M, Jones TA, Williams RD, Grigoriadis A, Vassal G, Workman P, Sheer D, Reis RM, Pearson AD, Hargrave D, Jones C - PLoS ONE (2009)

Bottom Line: All lines proliferate as adherent monolayers and express glial markers.Copy number profiling revealed complex genomes including amplification and deletions of genes known to be pivotal in core glioblastoma signalling pathways.Expression profiling identified 93 differentially expressed genes which were able to distinguish between the adult and paediatric high grade cell lines, including a number of kinases and co-ordinated sets of genes associated with DNA integrity and the immune response.

View Article: PubMed Central - PubMed

Affiliation: Paediatric Oncology, The Institute of Cancer Research, Sutton, United Kingdom.

ABSTRACT

Background: Although paediatric high grade gliomas resemble their adult counterparts in many ways, there appear to be distinct clinical and biological differences. One important factor hampering the development of new targeted therapies is the relative lack of cell lines derived from childhood glioma patients, as it is unclear whether the well-established adult lines commonly used are representative of the underlying molecular genetics of childhood tumours. We have carried out a detailed molecular and phenotypic characterisation of a series of paediatric high grade glioma cell lines in comparison to routinely used adult lines.

Principal findings: All lines proliferate as adherent monolayers and express glial markers. Copy number profiling revealed complex genomes including amplification and deletions of genes known to be pivotal in core glioblastoma signalling pathways. Expression profiling identified 93 differentially expressed genes which were able to distinguish between the adult and paediatric high grade cell lines, including a number of kinases and co-ordinated sets of genes associated with DNA integrity and the immune response.

Significance: These data demonstrate that glioma cell lines derived from paediatric patients show key molecular differences to those from adults, some of which are well known, whilst others may provide novel targets for evaluation in primary tumours. We thus provide the rationale and demonstrate the practicability of using paediatric glioma cell lines for preclinical and mechanistic studies.

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Related in: MedlinePlus

Immunophenotyping of paediatric glioma cell lines.All lines were grown as monolayers and stained with a variety of glial and stem cell markers including glial fibrillary acidic protein (GFAP), S100, vimentin, synaptophysin, nestin and CD133. H&E – haematoxylin and eosin. All images original magnification ×400.
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pone-0005209-g001: Immunophenotyping of paediatric glioma cell lines.All lines were grown as monolayers and stained with a variety of glial and stem cell markers including glial fibrillary acidic protein (GFAP), S100, vimentin, synaptophysin, nestin and CD133. H&E – haematoxylin and eosin. All images original magnification ×400.

Mentions: All paediatric glioma cell lines were derived from astrocytomas of differing grades arising in patients aged 3–16 years old. They were established in vitro as monolayer cultures, with doubling times of between 24–48 hours, and demonstrated a mixture of stellate and bipolar morphologies, with some cells of polygonal, cuboidal or flattened appearance (Table 1). The astrocytic nature of the cells was confirmed in culture by immunohistochemistry with a range of glial markers (Figure 1). All cells showed some degree of GFAP-positivity, although SF188 and Res259 were only weakly expressing. All cells were strongly positive for S100 and vimentin, with the exception of UW479, which was vimentin negative. There was little synaptophysin expression, save for the occasional isolated cell in SF188 and KNS42 cultures. There was a surprisingly high level of expression of stem cell markers in the paediatric glioblastoma lines, with both SF188 and KNS42 strongly positive for nestin, and containing 7% and 4% CD133-positive cells respectively.


Molecular and phenotypic characterisation of paediatric glioma cell lines as models for preclinical drug development.

Bax DA, Little SE, Gaspar N, Perryman L, Marshall L, Viana-Pereira M, Jones TA, Williams RD, Grigoriadis A, Vassal G, Workman P, Sheer D, Reis RM, Pearson AD, Hargrave D, Jones C - PLoS ONE (2009)

Immunophenotyping of paediatric glioma cell lines.All lines were grown as monolayers and stained with a variety of glial and stem cell markers including glial fibrillary acidic protein (GFAP), S100, vimentin, synaptophysin, nestin and CD133. H&E – haematoxylin and eosin. All images original magnification ×400.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2666263&req=5

pone-0005209-g001: Immunophenotyping of paediatric glioma cell lines.All lines were grown as monolayers and stained with a variety of glial and stem cell markers including glial fibrillary acidic protein (GFAP), S100, vimentin, synaptophysin, nestin and CD133. H&E – haematoxylin and eosin. All images original magnification ×400.
Mentions: All paediatric glioma cell lines were derived from astrocytomas of differing grades arising in patients aged 3–16 years old. They were established in vitro as monolayer cultures, with doubling times of between 24–48 hours, and demonstrated a mixture of stellate and bipolar morphologies, with some cells of polygonal, cuboidal or flattened appearance (Table 1). The astrocytic nature of the cells was confirmed in culture by immunohistochemistry with a range of glial markers (Figure 1). All cells showed some degree of GFAP-positivity, although SF188 and Res259 were only weakly expressing. All cells were strongly positive for S100 and vimentin, with the exception of UW479, which was vimentin negative. There was little synaptophysin expression, save for the occasional isolated cell in SF188 and KNS42 cultures. There was a surprisingly high level of expression of stem cell markers in the paediatric glioblastoma lines, with both SF188 and KNS42 strongly positive for nestin, and containing 7% and 4% CD133-positive cells respectively.

Bottom Line: All lines proliferate as adherent monolayers and express glial markers.Copy number profiling revealed complex genomes including amplification and deletions of genes known to be pivotal in core glioblastoma signalling pathways.Expression profiling identified 93 differentially expressed genes which were able to distinguish between the adult and paediatric high grade cell lines, including a number of kinases and co-ordinated sets of genes associated with DNA integrity and the immune response.

View Article: PubMed Central - PubMed

Affiliation: Paediatric Oncology, The Institute of Cancer Research, Sutton, United Kingdom.

ABSTRACT

Background: Although paediatric high grade gliomas resemble their adult counterparts in many ways, there appear to be distinct clinical and biological differences. One important factor hampering the development of new targeted therapies is the relative lack of cell lines derived from childhood glioma patients, as it is unclear whether the well-established adult lines commonly used are representative of the underlying molecular genetics of childhood tumours. We have carried out a detailed molecular and phenotypic characterisation of a series of paediatric high grade glioma cell lines in comparison to routinely used adult lines.

Principal findings: All lines proliferate as adherent monolayers and express glial markers. Copy number profiling revealed complex genomes including amplification and deletions of genes known to be pivotal in core glioblastoma signalling pathways. Expression profiling identified 93 differentially expressed genes which were able to distinguish between the adult and paediatric high grade cell lines, including a number of kinases and co-ordinated sets of genes associated with DNA integrity and the immune response.

Significance: These data demonstrate that glioma cell lines derived from paediatric patients show key molecular differences to those from adults, some of which are well known, whilst others may provide novel targets for evaluation in primary tumours. We thus provide the rationale and demonstrate the practicability of using paediatric glioma cell lines for preclinical and mechanistic studies.

Show MeSH
Related in: MedlinePlus