Modeling the Effects of Stroma on Clear Cell Renal Cell Carcinoma
Technical Report,01 Sep 2018,31 Aug 2019
MaineHealth Portland United States
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The year 1 project period was focused on 1. Obtaining regulatory compliance approvals from DoD, 2. Generating baseline data required for modeling clear cell renal cell carcinoma ccRCC, 3. Isolating primary ccRCC cells from patient samples. The regulatory approvals were issued within the first 3 months of the award, giving us 9 months to work on human tumor samples. We have successfully measured the rigidity of tumor samples and modified our silk scaffolding material to match. Two sequential window acquisition of all theoretical fragment ion spectra SWATH mass spectrometry experiments were run first a pilot to define parameters and identify problems, and then an experimental run on 7 tumors and neighboring healthy kidney tissue. This extensive dataset will be analyzed in the coming project period to define the extracellular matrix cocktail that we will use to scaffold primary tumor cells and fibroblasts. Primary tumor cells have been isolated and cryopreserved pending finalization of the scaffolding system. One challenge that has been identified is in the isolation of tumor associated fibroblasts in our hands these cells are not liberated using published dissociation techniques and we are in the process of empirically revising existing methods.
- Medicine and Medical Research