Implementation of a dynamic culture condition to the heterotypic 3D breast cancer model

Abstract

Cell culture system is used for a wide range of research and biotechnology production. Majority of in vitro cell studies are conducted as static, two dimensional (2D) dish culture system where cells grow in a monolayer. However, to better reflect the in vivo condition, three dimensional (3D) culture systems were introduced that allow investigating the cell–cell and cell–microenvironment interactions. In this work, the 3D breast cancer model was investigated. Previously, we developed a 3D breast cancer model that constituted of fibroblasts and breast cancer cells seeded on the silkworm silk scaffold. The dynamic culture condition that provides the medium flow and shear forces was implemented to the model. The dynamic conditions were compared to the static cultivation regarding its influence on the number of cells, their viability, scaffold penetration, and cells co‐localization. The implication of the dynamic condition to the 3D cultures resulted in a higher number and viability of the cells compared with the static 3D cultures. In contrast to the static culture condition, during the dynamic cultivation cells penetrated entirely and evenly the inner parts of the scaffold. Moreover, in coculture, the transitions like a ratio of fibroblast to the cancer cells, fibroblast morphology, and their localization were similar in both types of culture conditions, but they proceeded much faster during the dynamic cultivation. The implementation of dynamic culture condition shortened the time needed to establish the settle 3D breast cancer model. The established dynamic cancer model can be used to study tumor biology and drug screening.

Validation of a 3D perfused cell culture platform as a tool for humanised preclinical drug testing in breast cancer using established cell lines and patient-derived tissues

“…SY5Y neuroblastoma cells cultured as microtumours in Matrigel showed increased viability and volume after 7 days in perfused culture under monodirectional flow compared to static, with long term culture of up to 35 days possible [26]. Similarly, NIH3T3 fibroblasts or EMT6 BC cells growing alone or in co-culture on silk fibroin scaffolds showed higher cell viabilities than in static culture condition over 21 days, with perfusion generated by agitation [27]. We did not use any matrix in our experiments as cells appeared to grow well in the naked SeedEZ scaffold.…”

Proteinaceous Hydrogels for Bioengineering Advanced 3D Tumor Models

“…Recent studies have suggested that cells cultured under dynamic conditions show a higher cellular proliferation but still show the same morphology and behavior as cells cultured under static conditions. [ 163 ] Importantly, by using this approach, researchers were able to observe a limitation in oxygen and nutrients diffusion across the hydrogel, and at ≈150–200 µm depth an upregulation in the expression of HIF‐1 α was obtained. This was accompanied by an upregulation of vascular endothelial growth factor (VEGF)‐A expression, recreating the VEGF‐A expression promoted by HIF‐1 α similarly to that occurring in vivo, [ 164 ] suggesting the angiogenic potential of the tumor model, since VEGF‐A is involved in tumor angiogenesis.…”

From Donor to the Lab: A Fascinating Journey of Primary Cell Lines

“…Since the 1970s in the 20th century, many researchers have contributed to developing 3D cell culture, so cancer studies can be conducted on this model (Penderecka et al, 2020). The 3D culture shows significant improvements in studies concerning cell morphology, proliferation, response to stimuli, and drug metabolism.…”

“…Molecular mechanisms, such as gene expression, splicing, topology, and biochemistry of cells, are well preserved by contrast to 2D cultures that tend to interfere with gene expression (Kapałczyńska et al, 2016). The most common 3D models used for cancer research are scaffold-based cultures, multicellular spheroids, and matrix-embedded cultures (Penderecka et al, 2020).…”