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Cytometric analysis of cell suspension generated by cavitron ultrasonic surgical aspirator in pediatric brain tumors

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

Abstract

Purpose

The aim of this study was to test the possibility of using specimens obtained by a cavitron ultrasonic surgical aspirator (CUSA) in flow and mass cytometry investigations of pediatric brain tumors.

Methods

CUSA specimens obtained from 19 pediatric patients with brain tumors were investigated. Flow and mass cytometry methods were applied to analyze the composition of material collected using the CUSA. Cell suspensions were prepared from CUSA aspirates. Then sample viability was assessed by conventional flow cytometry and subsequently stained with a panel of 31 metal-labeled antibodies.

Results

Viability assessment was performed using conventional flow cytometry. Viability of cells in the acquired samples was below 50% in 16 of 19 cases. A mass cytometry investigation and subsequent analysis enabled us to discriminate brain tumor cells from contaminating leukocytes, whose proportions varied across the specimens. The addition of the viability marker cisplatin directly into the mass cytometry panel gave the means to selecting viable cells only for subsequent analyses. The proportion of non-viable cells was higher among tumor cells compared leukocytes.

Conclusions

When the analysis of the tumor cell immunophenotype is performed with markers for determining viability, the expression of the investigated markers can be evaluated. Suitable markers can be selected by high-throughput methods, such as mass cytometry, and those that are diagnostically relevant can be investigated using flow cytometry, which is more flexible in terms of time.

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Acknowledgements

This study was supported by Ministry of Health of the Czech Republic, grant nr. 15-26588A; Charles University (UNCE 204012); Research infrastructure was supported by Ministry of Education, Youth and Sports NPU I nr. LO1604; The CyTOF2 instrument was supported by EU-Prague project CZ.2.16/3.1.00/21540. Work of CyTOF2 operator Daniel Thürner is highly appreciated.

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Authors and Affiliations

  1. CLIP - Childhood Leukaemia Investigation Prague, Prague, Czech Republic

    Martina Vaskova, Daniela Kuzilkova, Tomas Kalina & Ondrej Hrusak

  2. Department of Paediatric Haematology and Oncology, Second Faculty of Medicine, Charles University and Motol University Hospital, Prague, Czech Republic

    Martina Vaskova, Daniela Kuzilkova, Ales Vicha, Jan Hrabeta, Tomas Kalina, Jan Stary & Ondrej Hrusak

  3. Department of Neurosurgery, Second Faculty of Medicine, Charles University and Motol University Hospital, Prague, Czech Republic

    Michal Tichy & Petr Liby

  4. Department of Pathology and Molecular Medicine, Second Faculty of Medicine, Charles University and Motol University Hospital, Prague, Czech Republic

    Josef Zamecnik

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  1. Martina Vaskova
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Correspondence to Martina Vaskova.

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Vaskova, M., Tichy, M., Zamecnik, J. et al. Cytometric analysis of cell suspension generated by cavitron ultrasonic surgical aspirator in pediatric brain tumors. J Neurooncol 143, 15–25 (2019). https://doi.org/10.1007/s11060-019-03135-w

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  • DOI: https://doi.org/10.1007/s11060-019-03135-w

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