Screening of additive manufactured scaffolds designs for triple negative breast cancer 3D cell culture and stem-like expansion

Breast cancer stem cells (BCSCs) are tumor-initiating cells responsible for metastasis and tumor reappearance, but their research is limited by the impossibility to cultivate them in a monolayer culture. Scaffolds are three-dimensional (3D) cell culture systems which avoid problems related with culturing BCSC. However, a standardized scaffold for enhancing a BCSC population is still an open issue. The main aim of this study is to establish a suitable poly (lactic acid) (PLA) scaffold which will produce BCSC enrichment, thus allowing them to be studied. Different 3D printing parameters were analyzed using Taguchi experimental design methods. Several PLA scaffold architectures were manufactured using a Fused Filament Fabrication (FFF) 3D printer. They were then evaluated by cell proliferation assay and the configurations with the highest growth rates were subjected to BCSC quantification by ALDH activity. The design SS1 (0.2 mm layer height, 70% infill density, Zigzag infill pattern, 45° infill direction, and 100% flow) obtained the highest proliferation rate and was capable of enhancing a ALDH+ cell population compared to 2D cell culture. In conclusion, the data obtained endorse the PLA porous scaffold as useful for culturing breast cancer cells in a microenvironment similar to in vivo and increasing the numbers of BCSCsThe authors acknowledge the financial support from the Ministry of Economy and Competitiveness (MINECO), Spain for the PhD scholarship and grants from DPI2016-77156-R and RYC-2014-15581, the financial support from the University of Girona (Spain) MPCUdG2016/036 and the support of the Catalan Government (2017SGR00385). This work was also partially supported by the grants from the Fundación Ramón Areces (Spain) and the Instituto de Salud Carlos III (Spain) (PI1400329