In Vivo Observation of Endothelial Cell-Assisted Vascularization in Pancreatic Cancer Xenograft Engineering

Abstract

In this study, for better understanding of patient-derived xenograft (PDX) generation, angiogenic characteristics during PDX cancerous tissue generation was investigated with different initial cell seeding conditions in the hydrogel. We monitored the angiogenic changes during the formation of in vivo cancer cell line xenografts induced by endothelial cells. Our in vivo cancer tissue formation system was designed with the assistance of tissue engineering technology to mimic patient-derived xenograft formation. Endothelial cells and MIA PaCa-2 pancreatic carcinoma cells were encapsulated in fibrin gel at different mixing configurations and subcutaneously implanted into nude mice. To investigate the effect of the initial cancerous cell distribution in the fibrin gel, MIA PaCa-2 cells were encapsulated as a homogeneous cell distribution or as a cell aggregate, with endothelial cells homogeneously distributed in the fibrin gel. Histological observation of the explanted tissues after different implantation periods revealed three different stages: isolated vascular tubes, leaky blood vessels, and mature cancerous tissue formation. The in vivo engineered cancerous tissues had leaky blood vessels with low expression of the vascular tight junction marker CD31. Under our experimental conditions, complex cancer-like tissue formation was most successful when tumorous cells and endothelial cells were homogeneously mixed in the fibrin gel. The present study implies that tumorous xenograft tissue formation can be achieved with a low number of initial cells and that effective vascularization conditions can be attained with a limited volume of patient-derived cancer tissue. Endothelial cellassisted vascularization can be a potent choice for the effective development of vascularized cancerous tissues for studying patient-derived xenografts, cancer angiogenesis, cancer metastasis, and anticancer drugs.

Combination of Irreversible Electroporation and STING Agonist for Effective Cancer Immunotherapy

“…Since 2D cell culture does not represent in vivo 3D tumor tissue, it is difficult to evaluate the effect of treatment with IRE accurately. To mimic in vivo conditions and evaluate cell death by IRE, an in vitro alginate-gelatin 3D-cell culture model (Figure 2b) was prepared and applied with IRE with a 3 mm two-needle array [41][42][43]. Live and dead assays were performed 24 h after applying 8, 40, and 80 pulses at 1000 V, which are the same conditions as the 2D experiment.…”

Polymer nanoparticle-assisted chemotherapy of pancreatic cancer

“…46 In addition, tumor blood vessels are immature and leaky, with low expression of the vascular tight junction marker CD31. 47 Furthermore, the immature vasculatures of cancer tissue are disordered, with small pore sizes. 48 Many factors can lead to changes of the oxygen level in the TME.…”