| Cancer is an extremely heterogeneous disease and the key to understanding and interpreting these differences lies in the unique molecular characteristics driving the disease. Using a micro-NMR detection system, we have previously found a biomarker signature that accurately diagnosed malignant cancer in patients using fine needle aspirate (FNAs) samples, which are less invasive than core biopsies but have limited clinical use due to its small sample size. To facilitate the use of FNAs for molecular diagnostics, we have developed a microfluidic platform for the dissociation of tumor samples into single cells exclusively using hydrodynamic forces for mechanical disruption. The dissociation chamber design was motivated by the goal of disrupting tissue while maintaining structural integrity of cells. Tumor models of increasing complexity were used to characterize the device. Concentration limits, ideal flow rates, cell viability, and biomarker integrity were evaluated. Tests were performed on both trypsin-EDTA treated cells and our in vitro models of tumor sheets and spheroids, which showed similar expression levels of propidium iodide and EpCAM in trypsin-EDTA treated samples. Single cell yields increased with increasing passes through the device for both in vitro models. A separate cell filter device was used and tested that yielded over 80% single cell recovery. |
