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Glioma cells showing IDH1 mutation cannot be propagated in standard cell culture conditions.

Piaskowski S, Bienkowski M, Stoczynska-Fidelus E, Stawski R, Sieruta M, Szybka M, Papierz W, Wolanczyk M, Jaskolski DJ, Liberski PP, Rieske P - Br. J. Cancer (2011)

Bottom Line: For example, glioblastoma cell lines presenting EGFR amplification cannot be established.IDH1 sequencing and loss of heterozygosity analysis was performed for 15 surgery samples of astrocytoma and early and late passages of cells derived from those and for 11 archival samples.Moreover, a new model for culturing glioma cells in vitro should be designed since the current one does not provide conditions corresponding to in vivo growth.

View Article: PubMed Central - PubMed

Affiliation: Department of Molecular Pathology and Neuropathology, Medical University of Lodz, Czechoslowacka 8/10 Street, Lodz 92-216, Poland.

ABSTRACT

Background: It has recently been reported by several sources that original (i.e., present in vivo) glioma cell phenotypes or genotypes cannot be maintained in vitro. For example, glioblastoma cell lines presenting EGFR amplification cannot be established.

Methods and results: IDH1 sequencing and loss of heterozygosity analysis was performed for 15 surgery samples of astrocytoma and early and late passages of cells derived from those and for 11 archival samples. We were not able to culture tumour cells presenting IDH1 mutations originating from currently proceeded 10 tumours; the same results were observed in 7 samples of archival material.

Conclusion: The IDH1 mutation is expected to be almost mutually exclusive with EGFR amplification, so glioma cells with IDH1 mutations seem to represent a new group of tumour cells, which cannot be readily analysed in vitro because of their elimination. The reasons for this intriguing phenomenon should be investigated since its understanding can help to define a new therapeutic approach based on simulating in vivo conditions, responsible for tumour cells elimination in vitro. Moreover, a new model for culturing glioma cells in vitro should be designed since the current one does not provide conditions corresponding to in vivo growth.

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Related in: MedlinePlus

IDH1 gene sequencing analysis of two astrocytic tumours (G59 and G60). The analysis of DNA from frozen samples shows a heterozygous mutation; both mutated and wild-type nucleotides are detected (1). The analysis of the second passage of corresponding culture cells shows the lack (G59), or almost invisible band (G60) of the mutated nucleotide (2). Mutated nucleotides are marked with arrows.
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fig1: IDH1 gene sequencing analysis of two astrocytic tumours (G59 and G60). The analysis of DNA from frozen samples shows a heterozygous mutation; both mutated and wild-type nucleotides are detected (1). The analysis of the second passage of corresponding culture cells shows the lack (G59), or almost invisible band (G60) of the mutated nucleotide (2). Mutated nucleotides are marked with arrows.

Mentions: DNA sequencing revealed IDH1 mutations in 10 surgical specimens of astrocytic tumours coming from 15 patients. The mutated template was no longer detectable in adherent cells grown in vitro (in four cases after the first passage and in six cases after the second passage) derived from the tumour samples with IDH1 mutations (Figure 1). Free floating cells (absorbing Trypan blue) from the medium showed IDH1 mutation after the first and the second passage. Our previous investigations showed that none of the tumours presented EGFR amplification, and three showed a mutation of TP53.


Glioma cells showing IDH1 mutation cannot be propagated in standard cell culture conditions.

Piaskowski S, Bienkowski M, Stoczynska-Fidelus E, Stawski R, Sieruta M, Szybka M, Papierz W, Wolanczyk M, Jaskolski DJ, Liberski PP, Rieske P - Br. J. Cancer (2011)

IDH1 gene sequencing analysis of two astrocytic tumours (G59 and G60). The analysis of DNA from frozen samples shows a heterozygous mutation; both mutated and wild-type nucleotides are detected (1). The analysis of the second passage of corresponding culture cells shows the lack (G59), or almost invisible band (G60) of the mutated nucleotide (2). Mutated nucleotides are marked with arrows.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3065269&req=5

fig1: IDH1 gene sequencing analysis of two astrocytic tumours (G59 and G60). The analysis of DNA from frozen samples shows a heterozygous mutation; both mutated and wild-type nucleotides are detected (1). The analysis of the second passage of corresponding culture cells shows the lack (G59), or almost invisible band (G60) of the mutated nucleotide (2). Mutated nucleotides are marked with arrows.
Mentions: DNA sequencing revealed IDH1 mutations in 10 surgical specimens of astrocytic tumours coming from 15 patients. The mutated template was no longer detectable in adherent cells grown in vitro (in four cases after the first passage and in six cases after the second passage) derived from the tumour samples with IDH1 mutations (Figure 1). Free floating cells (absorbing Trypan blue) from the medium showed IDH1 mutation after the first and the second passage. Our previous investigations showed that none of the tumours presented EGFR amplification, and three showed a mutation of TP53.

Bottom Line: For example, glioblastoma cell lines presenting EGFR amplification cannot be established.IDH1 sequencing and loss of heterozygosity analysis was performed for 15 surgery samples of astrocytoma and early and late passages of cells derived from those and for 11 archival samples.Moreover, a new model for culturing glioma cells in vitro should be designed since the current one does not provide conditions corresponding to in vivo growth.

View Article: PubMed Central - PubMed

Affiliation: Department of Molecular Pathology and Neuropathology, Medical University of Lodz, Czechoslowacka 8/10 Street, Lodz 92-216, Poland.

ABSTRACT

Background: It has recently been reported by several sources that original (i.e., present in vivo) glioma cell phenotypes or genotypes cannot be maintained in vitro. For example, glioblastoma cell lines presenting EGFR amplification cannot be established.

Methods and results: IDH1 sequencing and loss of heterozygosity analysis was performed for 15 surgery samples of astrocytoma and early and late passages of cells derived from those and for 11 archival samples. We were not able to culture tumour cells presenting IDH1 mutations originating from currently proceeded 10 tumours; the same results were observed in 7 samples of archival material.

Conclusion: The IDH1 mutation is expected to be almost mutually exclusive with EGFR amplification, so glioma cells with IDH1 mutations seem to represent a new group of tumour cells, which cannot be readily analysed in vitro because of their elimination. The reasons for this intriguing phenomenon should be investigated since its understanding can help to define a new therapeutic approach based on simulating in vivo conditions, responsible for tumour cells elimination in vitro. Moreover, a new model for culturing glioma cells in vitro should be designed since the current one does not provide conditions corresponding to in vivo growth.

Show MeSH
Related in: MedlinePlus