Establishment of a Pre-vascularized 3D Lung Cancer Model in Fibrin Gel—Influence of Hypoxia and Cancer-Specific Therapeutics

Abstract

Lung cancer is the most frequently diagnosed cancer worldwide and the one that causes the highest mortality. In order to understand the disease and to develop new treatments, in vitro human lung cancer model systems which imitate the physiological conditions is of high significance. In this study, a human 3D lung cancer model was established that features the organization of a tumor with focus on tumor angiogenesis. Vascular networks were formed by co-culture of human umbilical vein endothelial cells and adipose tissue-derived mesenchymal stem cells (ASC) for 14 days in fibrin. A part of the pre-vascularized fibrin gel was replaced by fibrin gel containing lung cancer cells (A549) to form tri-cultures. This 3D cancer model system was cultured under different culture conditions and its behaviour after treatment with different concentrations of tumor-specific therapeutics was evaluated. The evaluation was performed by measurement of metabolic activity, viability, quantification of two-photon laser scanning microscopy and measurement of the proangiogenic factor vascular endothelial growth factor in the supernatant. Hypoxic conditions promoted vascularization compared to normoxic cultured controls in co- and tri-cultures as shown by significantly increased vascular structures, longer structures with a higher area and volume, and secretion of vascular endothelial growth factor. Cancer cells also promoted vascularization. Treatment with 50 µM gefitinib or 50 nM paclitaxel decreased the vascularization significantly. VEGF secretion was only reduced after treatment with gefitinib, while in contrast secretion remained constant during medication with paclitaxel. The findings suggest that the herein described 3D lung cancer model provides a novel platform to investigate the angiogenic potential of cancer cells and its responses to therapeutics. Thus, it can serve as a promising approach for the development and patient-specific pre-selection of anticancer treatment.

In vitro vascularized immunocompetent patient-derived model to test cancer therapies

“…Their presence in the tumor culture not only supports the formation of intact cellular assemblies but also offers an opportunity to directly assess angiogenic properties in vitro 49 , 50 and the activity of antiangiogenic inhibitors. 51 We studied the typical morphology of Ad-MVs and their ability to enhance some vascular network formation within our PDT model ( Figure 4 A). On day 7 of the PDTs formation, the vascular network within the PDTs was characterized with UEA-1 lectin and CD31 staining.…”

Design, Synthesis and Biological Evaluation of 6-(Imidazo[1,2-a]pyridin-6-yl)quinazoline Derivatives as Anticancer Agents via PI3Kα Inhibition

“…Hence, it was widely applied in research fields including new drug screening, tumour cell system biology, stem cell research and functional tissue implantation [46][47][48]. Additionally, previous findings indicated that the phenotype of the 3D lung cancer tumour sphere in vitro is closer to that of real cancer tissue in vivo [49,50]. Thus, it is considered a reasonable method to evaluate the in vivo efficacy of active compounds in the early stages of new drug development [51].…”

Mimicking Molecular Pathways in the Design of Smart Hydrogels for the Design of Vascularized Engineered Tissues

“…In vitro, a hydrogel material can be created by mixing fibrinogen with thrombin and calcium ions [99][100][101]. Fibrin-based hydrogels, renowned for their ability to promote vasculogenesis, are commonly employed in various models of vascularization due to their ease of fabrication [99,100,[102][103][104][105]. Furthermore, fibrin can be recovered from blood for the creation of autologous hydrogel, which ensures that viable implants can be used therapeutic applications [106].…”