Morphology of Adherent Cells of the Line Vero Cultivated in a Three-Dimensional Environment inside a Microfluidic Device Differs from their Morphology when Cultivated in Monolayers

Abstract

Traditional in vitro culture models have significant limitations in mimicking important physiological interactions, such as cell-cell interactions, cell-extracellular matrix interactions, and the three-dimensional morphology of cells. In contrast, 3D culture models have the ability to replicate the natural three-dimensional environment of cells. Aims: The objective of this study is to evaluate the morphology of adherent Vero cells grown in two-dimensional (2D) and three-dimensional (3D) culture models. Methodology: For the 2D culture model, Vero cells were thawed and grown in culture flasks in nutrient-rich culture medium. As for the 3D culture, a collagen hydrogel solution was prepared to mimic the extracellular matrix and injected into the central microchamber of the microfluidic device along with the Vero cells. To compare the morphological differences between the two culture models, measurements of the shortest and longest axes of the cells were performed, and the proportion of the cell axes in the two types of culture was compared. Results: The results indicated that in both 2D and 3D cultures, the minor axis of the cells has similar sizes, being 106 ± 22.7 and 109.9 ± 35.8 µm, respectively. However, the major axis of the cells in 3D culture was significantly larger, compared to 2D, with values ​​of 154.8 ± 11.96 and 114.1 ± 6.25, respectively. Similarly, Vero cells had a higher proportion value, being 1.08 ± 0.11 and 1.48 ± 0.39, respectively for 2D and 3D cultures. Conclusion: We conclude that Vero cells in a 3D environment have a different morphology than cells cultured in 2D. One of the main differences is related to the size of the largest cell axis and consequently the proportion of the axes. The data suggest that in 3D cultures, cells are more elongated, with filopodia involved in cell-cell and cell-ECM interactions.