Article
Brain tumor tissue obtained by ultrasonic aspiration for primary cell culture and histopathological evaluation – Technical note
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Published: | June 4, 2012 |
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Objective: Ultrasonic aspiration has been widely adopted in the resection of tumors of the central nervous system. In the past, the tumor tissue fragments obtained have been discarded. To evaluate these fragments as possible sources of material for histopathological study and tissue culture, we compared the microscopic features and viability in tissue culture of CUSA tissue fragments and biopsies obtained by conventional methods.
Methods: Brain tumor tissue obtained by ultrasonic aspiration (CUSA EXcelTM, Integra Radionics, Inc.) into a simple suction trap during resection was processed for primary cell cultures. Samples were cleared by centrifugation, washed with DMEM (Invitrogen, Darmstadt, Germany) and digested with trypsin 30 minutes at 37°C for at least one time until no fragments could be explored. Cells were seeded in DMEM as monolayer cultures supplemented with 10% heat-inactivated fetal calf serum, 100 U/ml penicillin and 100 μg/ml streptomycin and 2 mM L-glutamine (all Invitrogen, Darmstadt, Germany). Further, cells were cultured in T75 tissue culture flasks (Greiner BioOne, Frickenhausen, Germany) at 37°C and 5% CO2 in a humidified atmosphere. Cell viability was measured by WST-1 Test and immunohistological evaluation was performed in the Institute of Neuropathology.
Results: Eight cases (one meningioma, two metastasis, and 5 gliomas) are presented to demonstrate that these tissue fragments retain good preservation of histological detail and tissue culture viability.
Conclusions: Brain tumor tissue available from a simple suction trap during resection with the CUSA ExcelTM (Integra Radionics, Inc.) is consistently greater than that obtained by standard biopsy methods. Utilization of this material may facilitate pathological interpretation by providing a more representative sample of tumor histology as well as providing an adequate and sterile source of material for tissue culture studies.