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Characterization of primary glioma cell lines derived from the patients according to 2016 CNS tumour WHO classification and comparison with their parental tumours

  • Laboratory Investigation
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Journal of Neuro-Oncology Aims and scope Submit manuscript

Abstract

Background

Gliomas represent about 80% of primary brain tumours and about 30% of malignant ones, which today don’t have a resolution therapy because of their variability. A valid model for the study of new personalized therapies can be represented by primary cultures from patient’s tumour biopsies.

Methods

In this study we consider 12 novel cell lines established from patients’ gliomas and immunohistochemically and molecularly characterized according to the newly updated 2016 CNS Tumour WHO classification.

Results

Eight of these lines were glioblastoma cells, two grade III glioma cells (anaplastic astrocytoma and oligo astrocytoma) and two low grade glioma cells (grade II astrocytoma and oligodendroglioma). All cell lines were analysed by immunohistochemistry for specific glioma markers, respectively VIMENTIN, GFAP, IDH1R132, and ATRX. The methylation status of the MGMT gene promoter was also determined in all lines. The comparison of the immunohistochemical characteristics and of the MGMT methylation status of the lines with the tissues of origin shows that the cells in culture maintain the same characteristics.

Conclusions

Human cancer cell lines represent a support in the knowledge of tumour biology and in drug discovery through its facile experimental manipulation.

Trial registration

NCT 04180046.

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Data availability

The data that support the findings of this study are available from the authors upon reasonable request.

Abbreviations

GBM:

Glioblastoma

TMZ:

Temozolomide

BBB:

Blood–brain barrier

IDH:

Isocitrate dehydrogenase

MGMT:

O6-methylguanine–DNA methyltransferase

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Funding

This work has been supported by Italian Ministry of Health with Ricerca Corrente.

Author information

Authors and Affiliations

  1. Via Atinense 18, IRCCS Istituto Neurologico Mediterraneo NEUROMED, 86077, Pozzilli, IS, Italy

    Maria Antonietta Oliva, Sabrina Staffieri, Salvatore Castaldo, Felice Giangaspero, Vincenzo Esposito & Antonietta Arcella

  2. University of Rome La Sapienza, Rome, Italy

    Felice Giangaspero & Vincenzo Esposito

Authors
  1. Maria Antonietta Oliva
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  2. Sabrina Staffieri
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  3. Salvatore Castaldo
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  4. Felice Giangaspero
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  5. Vincenzo Esposito
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  6. Antonietta Arcella
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Contributions

MAO performed immunofluorescence, immunohistochemistry and proliferation assay; SS performed MGMT methylation specific PCR and DNA sequencing; SC contributed to the expansion of cell cultures; VE collected and provided glioma biopsies; FG performed the histological diagnosis of gliomas and contributed to the discussion of the data; AA performed the molecular analysis of gliomas, coordinated this study, prepared primary cell cultures from biopsies and wrote the paper.

Corresponding author

Correspondence to Antonietta Arcella.

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Conflict of interest

The authors declare that they have no competing interests.

Consent for submission

The work described here has not been published before and is not under consideration for publication elsewhere. The authors declare that they have no conflict of interest. All authors have approved the manuscript.

Ethical approval

This study was approved by Neuromed Ethics Committee on 20 February 2020 and registered on Clinical Trial.gov with identification number NCT 04180046.

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Supplementary Information

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11060_2020_3673_MOESM1_ESM.tif

Figure S1 Primary cell culture and corresponding tumour tissue FL : low grade human glioma (grade II oligodendroglioma) IDH1R132 mutant. A) Cells and tissues express glioma markers VIMENTIN, GFAP, wild type ATRX and IDH1R132H mutation. B) Electropherogram shows cDNA Sequencing of IDH1 in primary cell line (lower panel) and DNA Sequencing of IDH1 in corresponding tumor tissue (upper panel). C) Representative electropherogram of Loss of heterozygosity analysis on 1p chromosome; it shows a deletion at locus D1S508 both in cells and corresponding patient's tissue respect to the patient's blood (used as control wild type). Supplementary file1 (TIF 55990 KB)

11060_2020_3673_MOESM2_ESM.tif

Figure S2 Doubling time of primary glioblastoma cell lines (A) and low grade glioma cell lines (B). Low grade glioma cells (MI, FL) proliferate slowly than glioblastoma cells (PAP, CL, NULU, COGI). Growth curves of low grade glioma cells IDH1 mutant treated with TMZ (C) Both glioma cell lines (MI, FL) were responsive to TMZ treatment, expecially at the higher concentration used (50 μM). Western blot analysis of IDH1R132H mutation in low grade glioma cells (D). Western blot of MI and FL protein’s lysates showed a 46kDa band corresponding to the mutated protein in both cell lines analysed. Supplementary file2 (TIF 20839 KB)

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Oliva, M.A., Staffieri, S., Castaldo, S. et al. Characterization of primary glioma cell lines derived from the patients according to 2016 CNS tumour WHO classification and comparison with their parental tumours. J Neurooncol 151, 123–133 (2021). https://doi.org/10.1007/s11060-020-03673-8

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  • DOI: https://doi.org/10.1007/s11060-020-03673-8

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