Abstract
A challenge in cancer research is developing reproducible, reliable, and practical models which can capture the complexity of cancer development and treatment. The development of functional precision medicine platforms is emerging as a promising strategy for improving pre-clinical drug testing and guiding clinical decisions. We have developed an organotypic brain slice culture (OBSC) technology composed of intact multicellular tissue which can be rapidly used for spatiotemporal drug response testing. OBSCs are reproducibly generated from Sprague-Dawley rat pups and used as living tissue substrates to culture treat different tumor cell lines and uncultured patient brain tumor resection tissue. We evaluated OBSC quality and reproducibility throughout the study by using a propidium iodide nuclear permeability assay. This technique enabled a broad dynamic range to distinguish between healthy and unhealthy slices. In these studies, the rat pup age at the time of generation had an impact on OBSC quality and quantity, indicating that OBSCs should be generated from eight-day-old pups. We also found that optimal OBSCs were generated using improved methods for brain dissection and OBSC culture conditions, establishing our robust, standardized procedure for OBSC generation. Following this optimization period, we continued to conduct quality control analysis with 6 OBSCs per batch for reproducibility. We also conducted immunohistochemistry analysis immediately after slicing and concluded that morphology of neurons in OBSCs remained unchanged between day 0 and 4. In addition, the activation of astrocytes attenuates by day 4 but persists in macrophages/microglia, suggesting that the myeloid cells can phagocytose dead cells and debris in OBSCs. In summary, these results indicate that the OBSC platform may be an effective model that accelerates preclinical drug testing and directs drug development towards clinical evaluation.