In vitro biological dosimeter modeling of the glioblastoma response to radiation delivered by the Gamma Knife

Abstract

The goal of this study was to develop an assay that makes possible the assessment of the glioma cell response to single-fraction high-dose Gamma Knife surgery. In this assay, the isolation of radioresistant cell subpopulations facilitates mechanistic studies of radioresistance. A tissue-equivalent paraffin phantom with an aperture capable of holding an Opticell cell culture cassette was developed for treatment with the Leksell Gamma Knife model C. A second apparatus, which the authors also created, uses the manufacturer-supplied polystyrene phantom, thereby allowing this assay to be performed in the Leksell Gamma Knife Perfexion. After treatment, the cells were morphologically assessed to determine their response to radiation treatment. Two specific parameters were used to determine radiosensitivity: 1) the diameter of the clearing zone, defined as the central region of cell death; and 2) the number of surviving colonies within this central high-dose clearing zone. Radioresistance was compared in 2 different cell lines from glioblastomas. The first cell line, ME, was established from a primary tumor before its treatment, and the second cell line, DIV, was established from a tumor that recurred after treatment with chemotherapy and fractionated radiotherapy. The ME cell line had the most robust response to radiosurgery, as characterized by a consistently larger clearing zone (28.33 ± 1.1 mm). In contrast, the clearing zone produced when the DIV cell line was used was 24.0 ± 1 mm, indicating an approximate response difference of 5 Gy. The mean number of surviving colonies within the clearing zone for the ME cell line was 1.33 ± 1 compared with that for the DIV cell line, which was 66.67 ± 2. The authors developed a biological dosimeter to model the response of cells from glioblastomas to single-fraction high-dose radiation. This system also allows the identification and isolation of radioresistant cells.