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  • Tissue Extraction and Preservation for Glioblastoma Specimens Used for Molecular Oncology and Personalized Therapy Studies

    Final Number:

    Liliana Soroceanu MD, PhD; Jimmin Chang PhD, RN; Maxim Sidorov; Alexandria M Ayala BS; Edie E. Zusman MD; Lawrence D. Dickinson MD

    Study Design:
    Laboratory Investigation

    Subject Category:

    Meeting: Congress of Neurological Surgeons 2017 Annual Meeting

    Introduction: The ability to establish cell cultures and animal models of disease that preserve the genomic profile and phenotype of the parental tumor, and can be used for in-vitro and in-vivo studies, has the potential to revolutionize development of new, individualized treatments for Glioblastoma. In this study, we assessed how the collection methodology and tissue preservation techniques, during intra-op, can be used to enhance the viability of tumor tissue for establishing primary cultures and patient derived xenografts (PDX) in immunocompromised mice.

    Methods: Fresh brain tumor tissue samples were obtained by conventional en-bloc resection technique and through use of a non-ablated piecemeal aspiration extraction system, with an automated tissue preservation module that flushes and chills the specimen as it is collected in a filter. Both specimens were then bathed in hypothermosol solution and kept cool during transport to laboratory for processing within 2 hours from surgical removal. Percentage of viable cells for primary cultures was determined using an automated cell counter. Matched specimens from the same patient were also interrogated using time course viability, and in-vivo tumorigenicity assays.

    Results: Comparison of en-bloc and aspiration extracted specimens indicated that aspiration resected samples can be effectively used for establishing cell cultures and PDX in vivo. Immediate flushing and chilling of the fresh tissues can increase viable cell count by at least 10%, but does not change long term cell viability or in-vivo tumorigenicty. Both en bloc and the aspiration resected samples were able to induce tumor growth in vivo, in an intracranial PDX model.

    Conclusions: Aspiration resection is currently the safest, most precise and least disruptive method for resecting glioblastoma. We have demonstrated that specimens collected using such tools, as compared to en-bloc excision, consistently yield robust cell cultures that can be used in generating PDX models for research, testing of individualized therapy, and possibly vaccine development.

    Patient Care: Aspiration resected tissue specimens can be used effectively for genomic analysis, and to produce cell cultures for use in in-vivo and in-vitro testing for molecular oncology and personalized therapy studies. Aspiration resection tools can be used to effectively collect specimen from the peripheral region of the tumor where latest genetic mutation information can be obtained for effective diagnosis and treatment.

    Learning Objectives: By the conclusion of this session, participants should be able to: (1) Describe how aspiration resected tissue specimens can be used effectively to produce cell cultures compared to en-bloc samples. Both specimens can be used for genomic analyses, in-vitro and in-vivo testing. (2) Describe how individual patient-derived cell cultures and mouse avatars can be used to identify personalized glioblastoma treatment. (3) Describe how automated tissue collection with immediate chilling and flushing of the fresh tumor tissue improves tissue preservation and increases cell viability.

    References: Inda M, Bonavia R, Seoane J. Glioblastoma Multiforme: A Look Inside Its Heterogeneous Nature. Cancers, 6:225-239, 2014. Schroeteler J, Reeker R, Suero Molina E, Brokinkel B, Holling M, Grauer OM, Senner V, Stummer W, Ewelt C. Glioma tissue obtained by modern ultrasonic aspiration with a simple sterile suction trap for primary cell culture and pathological evaluation. European Surgical Research, 53(1-4): 37-42, 2014. McLaughlin N, Ditzel Filho LF, Prevedello DM, Kelly DF, Carrau RL and Kassam AB. Side-Cutting Aspiration Device for Endoscopic and Microscopic Tumor Removal. Journal of Neurological Surgery: Skull Base. 73:11, 2012. Carlson BL, Pokorny JL, Schroeder MA, Sarkaria JN. Establishment, maintenance and in vitro and in vivo applications of primary human glioblastoma multiforme (GBM) xenograft models for translational biology studies and drug discovery. Current Protocol in Pharmacology, 52(14): 1-14, doi:10.1002/ 0471141755.ph1416s52, March 2011. Blackie R, Gordon A. Histological Appearances of Intracranial Biopsies Obtained using the Cavitron Ultrasonic Surgical Aspirator. Journal of Clinical Pathology, 37:1101-1104, 1984.

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