OP221. Application of a microfluidics system for evaluating the response of Glioblastoma tissues to radiotherapy and chemotherapy

摘要

Cancer is a highly heterogeneous disease characterised by multiple genetic lesions and aberrations. It is becoming increasingly clear that the molecular variations that exist within tumours predict prognosis and response to treatment. One way to overcome this is to optimise treatment(s) pre-clinically by studying each tumour on an individual basis. The use of patient derived tumour xenografts (in mice) as a disease avatar to aid in the selection of appropriate chemotherapeutic agents is a theoretical solution but the financial burden of this approach makes it unrealistic. Microfluidics is a fast growing area of research that allows experimentation with mimicry of natural conditions. Fluid flow through micro-devices takes place at a submilliliter scale, where viscous as opposed to inertial forces dictate flow; this equates to laminar flow and hence diffusion becomes the predominant form of cellular interactions. The dynamics of fluid at the micro-scale has been exploited with a variety of biological applications, such as protein crystallization, PCR, single cell analysis, chemotaxis, and evolutionary biology. In its simplest form a microfluidic environment allows the continuous influx and efflux of nutrients into a cell, cell culture or tissue but there are devices that have been created that mimic physiological organs such as the lung, heart or complex interplays between such organs. With regards to the study of cancer, microfluidic devices have been used to maintain biopsy samples of human cancer tissues for between 7–10 days in vitro. During this time period, the effects of chemotherapy and radiotherapy on the tissues have been tested and active cell death from the toxicity of the drugs and radiation has been confirmed. Though the literature boasts studies of head and neck, prostate and ovarian cancers within microfluidic devices, to date, there are relatively few studies on the application of microfluidics in Glioblastoma. Here, we present the preliminary results of our experience with human glioblastoma tissue samples maintained on a Polydimethylsiloxane (PDMS)/glass microfluidic platform. The design of the device is similar to that used to maintain viable head and neck squamous cell carcinoma tissues for up to 7 days and test their response to chemo-radiotherapy. We present our study protocol including tissue handling, culture (with continuous influx and efflux of media for 5 days) and the results of viability testing, including LDH and MTS assays which assess apoptotic cell death and cell proliferation respectively. We also compare histology and immunohistochemistry of fresh biopsies retrieved from the patients and those which were maintained within the microfluidic device for 5 days to prove the method does not affect tissue architecture. As far as we are aware, this is the first time that human glioblastoma tissue have been cultured ex vivo within a microfluidic device and we postulate that this modality is a practical and cost effective option for personalising drug treatments for this group of patients as well as an option for testing novel therapies. We believe that this work will provide a new platform for studying the biology of brain tumours.