Developing and characterizing stroma-enriched 3D $in~vitro$ tumor models for imaging and drug delivery applications
RWTH Publications (RWTH Aachen)
Abstract
Due to the high cost and time consumption, various ethical concerns, and poor predictive outcomes for human clinical trials, there is a strong interest in finding alternatives to animal testing. Recent advancements in the use of multicellular tumor spheroids and sophisticated tumor-on-chip models have shown comparable results to in vivo animal models.Taking this together, this thesis has explored development of various in vitro preclinical cancer models, ranging from 2D co-cultures, 3D tumor spheroids to dynamic cultivation systems for bridging the gap between simple 2D cultures and in vivo tumor models. It evaluates nanoparticle uptake, cytotoxicity, combination therapies, and tumor-stroma interactions driven by distinct CAF subtypes. Cytotoxicity assessment on 3D hetero-spheroids was conducted by monitoring changes in spheroid size, performing cell viability assays, staining for cell death markers, and evaluating drug distribution. Further, CAF subtypes were generated in vitro via cytokine stimulation, and the effect of specific CAF subtype and cytokine stimulation on tumor spheroid characteristics were systematically analyzed during long-term cultivation. The study further assessed the robustness of dynamically cultivated mesoscopic bioreactors by growing fibrotic and angiogenic patient-derived tumor spheroids and comparing outcomes with xenograft mouse models. Additionally, these bioreactors were also employed to investigate CAF-driven modulation of tumors and their microenvironments, measuring tumor spheroid growth, vascular network assembly and, fibrotic activity through 2D fluorescence reflectance imaging using a PDGFR-β-specific BIPPB probe. This thesis represents the first application of mesoscopic bioreactors for molecular imaging. Overall, the thesis provides valuable insights into tumor-stromal interactions and demonstrates the value of in vitro systems for therapy screening and molecular imaging in preclinical cancer research.
Authors 1
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Affiliation as printed
RWTH Aachen