Characterisation of 3D Bioprinted Human Breast Cancer Model for In Vitro Drug and Metabolic Targeting

Abstract

Monolayer cultures, the less standard three-dimensional (3D) culturing systems, and xenografts are the main tools used in current basic and drug development studies of cancer research. The aim of biofabrication is to design and construct a more representative in vivo 3D environment, replacing two-dimensional (2D) cell cultures. Here, we aim to provide a complex comparative analysis of 2D and 3D spheroid culturing, and 3D bioprinted and xenografted breast cancer models. We established a protocol to produce alginate-based hydrogel bioink for 3D bioprinting and the long-term culturing of tumour cells in vitro. Cell proliferation and tumourigenicity were assessed with various tests. Additionally, the results of rapamycin, doxycycline and doxorubicin monotreatments and combinations were also compared. The sensitivity and protein expression profile of 3D bioprinted tissue-mimetic scaffolds showed the highest similarity to the less drug-sensitive xenograft models. Several metabolic protein expressions were examined, and the in situ tissue heterogeneity representing the characteristics of human breast cancers was also verified in 3D bioprinted and cultured tissue-mimetic structures. Our results provide additional steps in the direction of representing in vivo 3D situations in in vitro studies. Future use of these models could help to reduce the number of animal experiments and increase the success rate of clinical phase trials.

3D bioprinting and the revolution in experimental cancer model systems—A review of developing new models and experiences with in vitro 3D bioprinted breast cancer tissue-mimetic structures

“…were used to construct 3D bioprinted models for performing a metabolic comparison with the same cell lines maintained in different culture systems. As we previously described, ZR75.1 luminal B breast cancer cells formed lumens after bioprinting and some-day maintenance ( 103 ). In this presented brief study, the continuous and significant growth activity of 3D bioprinted tissue-mimetic structures was detected by different proliferation tests or cell number analyses from 3 to 5 days after bioprinting to ∼3 weeks.…”

“…The printing conditions were the following: radius and height (2.5–5 mm, 0.5–1 mm); interlayer angle (90°); the distance of infill (1.5 µm); printing speed (10 mm/s); needle diameter and height (110 µm for cell-free-gel and 50–50 µm for cell-gel); pressure (400 kPa for cell-free-gel and 20 kPa for cell-gel). The scaffolds were post-processed by CaCl 2 crosslinking (200 mM, 2 min) and washed twice then maintained in culture media ( 103 ).…”

Shape Fidelity Evaluation of Alginate-Based Hydrogels through Extrusion-Based Bioprinting

“…The filament deformation test is based on previous works to assess the mid-point deflection of a hanged filament [ 11 , 48 ]. A custom platform of 1.6 × 2.75 × 4 mm (length, width, and height) with pillars placed at 1, 2, 4, 8, and 16 mm along the platform was designed in SolidWorks (Dassault Systèmes SolidWorks Corp., Waltham, MA, USA).…”

“…The three-dimensional (3D) bioprinting technology refers to the layer-by-layer patterning of cell-laden bioink(s) in a predefined structural design [ 1 , 2 , 3 ]. Applications of 3D bioprinting range from microfluidics, organ-on-chip technologies, and tissue engineering to real-sized organ implants [ 4 , 5 , 6 , 7 , 8 , 9 , 10 , 11 , 12 , 13 , 14 , 15 , 16 , 17 , 18 , 19 , 20 , 21 ]. Commonly used 3D bioprinting methods are laser-assisted bioprinting [ 22 ], stereolithography (SLA) [ 17 , 23 ], inkjet-based bioprinting [ 24 ], valve-based bioprinting [ 25 ], and extrusion-based bioprinting (EBB) [ 26 ].…”

Inhibition of Viral RNA-Dependent RNA Polymerases by Nucleoside Inhibitors: An Illustration of the Unity and Diversity of Mechanisms

“…Mammalian cells are traditionally cultured in two-dimensional monolayers for initial studies of virus infection and drug sensitivity screening. However, it has been increasingly acknowledged that a three-dimensional cell culture may better approximate metazoan tissues and organs and may be suitable for diverse studies such as “epithelial–mesenchymal transition”, drug uptake, and virus growth [ 88 , 89 , 90 , 91 , 92 ]. Many viruses show stringent preference for either the apical or the basolateral membrane of the cell [ 93 , 94 ], which are better studied in 3D cultures.…”