Human Lung Cancer Cells Grown in an Ex Vivo 3D Lung Model Produce Matrix Metalloproteinases Not Produced in 2D Culture

Abstract

We compared the growth of human lung cancer cells in an ex vivo three-dimensional (3D) lung model and 2D culture to determine which better mimics lung cancer growth in patients. A549 cells were grown in an ex vivo 3D lung model and in 2D culture for 15 days. We measured the size and formation of tumor nodules and counted the cells after 15 days. We also stained the tissue/cells for Ki-67, and Caspase-3. We measured matrix metalloproteinase (MMP) levels in the conditioned media and in blood plasma from patients with adenocarcinoma of the lung. Organized tumor nodules with intact vascular space formed in the ex vivo 3D lung model but not in 2D culture. Proliferation and apoptosis were greater in the ex vivo 3D lung model compared to the 2D culture. After 15 days, there were significantly more cells in the 2D culture than the 3D model. MMP-1, MMP-9, and MMP-10 production were significantly greater in the ex vivo 3D lung model. There was no production of MMP-9 in the 2D culture. The patient samples contained MMP-1, MMP-2, MMP-9, and MMP-10. The human lung cancer cells grown on ex vivo 3D model form perfusable nodules that grow over time. It also produced MMPs that were not produced in 2D culture but seen in human lung cancer patients. The ex vivo 3D lung model may more closely mimic the biology of human lung cancer development than the 2D culture.

Three-Dimensional Cell Culture Systems and Their Applications in Drug Discovery and Cell-Based Biosensors

“…Much more expensive for large-scale studies and high throughput assays than traditional 2D culture 17 Some models produce spheroids that vary greatly in size, resulting in high variability within the same well/flask 17 3D models still lack vasculature which plays a vital role in tumor growth/ survival and drug delivery 74,75 the electrical behavior of the spheroid includes the effective extracellular resistance of the spheroid, R ext ; the effective intracellular resistance of the spheroid, R int ; the effective cell membrane capacitance of the spheroid, C mem ; the electrical behavior of the detection system, which includes the stray capacitance of the system, C s ; and the resistance of the culture medium, R med . 89 Kloss et al demonstrated that drug-induced apoptosis in spheroids displayed increased impedance magnitudes, resulting from an altered morphology of the outer cells, using the microcavity system.…”

Effects of Secretome from Dynamic 3D Cell Culture System onto Growth and Cytoprotection of Nasal Fibroblast

“…Previous studies have shown that 3D cultured cells produce more unique proteins than 2D cultured cells and also the common secreted proteins in 2DCM and 3DCM was found to be enriched in 3DCM (Mishra et al 2012;Tit-Oon et al 2014). Thus, an in-depth proteomic analysis of the secretome can reveal the identity and function of the unique proteins found in this study.…”

The role of cell lysis and matrix deposition in tumor growth modeling

“…Tumor cells can in fact be grown successfully in a decellularized ECM of an organ as shown in Mishra et al [43]. This allows mimicking the in vivo environment and has been done successfully for an ex vivo 3D lung model where it was possible to grow perfusable lung nodules [43]. The simulations are limited to the avascular stage.…”