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UBC Theses and Dissertations

Organ decellularization used as a novel approach to engineer three-dimensional urogenital tumor models Tan, Zheng


Cancer is currently the leading cause of death in Canada and is responsible for 30% of all deaths. With various treatments available nowadays, it is necessary to develop a three-dimensional (3D) model to mimic in vivo condition of patients’ disease in order to test drug efficacy prior to any actual treatments. My study is focusing specifically on kidney and bladder cancer. Therefore, a 3D, acellular organ-specific extracellular matrix (ECM) can be established as bioactive substrates with the long-range goal of bioengineering tumors for drug testing. Specifically, decellularization is the process by which cells are removed from the organ to produce the aforementioned matrix. It is crucial to achieve a tight balance between effective cell removal and retention of native ECM architecture. Henceforth, the decellularized matrix can be used as a template and be repopulated with normal human organ-specific cells or cancerous cells. With these approaches, a bio-matrix that ultimately serves as a cancer model for drug testing can be developed. In this master’s thesis project, I hypothesized that decellularization preserves organ microarchitecture and retains various matrix-bound growth factors necessary for cell homing. Afterward, tumor models can be established through recellularization of human cells onto the decellularized ECMs as a proof-of-principle drug-testing platform to predict treatment response. Two objectives were pursued to test this hypothesis. Under objective 1, I optimized and established decellularization protocols for both kidney and bladder. The protocols ensure complete cell removal and preservation of ECM microarchitectures. In addition, comprehensive protein-profiling of the decellularized kidney and bladder’s ECM was completed by performing proteomic analysis with liquid chromatography–mass spectrometry (LC-MS/MS). Under objective 2, recellularization protocols for both decellularized kidney and bladder ECM were established and optimized. As the decellularized kidney and bladder’s ECM was repopulated with normal human cells and cancerous cells, the proposed in vitro urogenital cancer model can be developed. In conclusion, a 3D in vitro urogenital cancer model can be developed from kidneys and bladders using protocols optimized under objective 1 & 2. This work establishes the model as proof –of principal and sets the foundation for the developments of personalized cancer treatments.

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