Differentiation capacity and maintenance of differentiated phenotypes of human mesenchymal stromal cells cultured on two distinct types of 3D polymeric scaffolds

Abstract

Many studies have shown the influence of soluble factors and material properties on the differentiation capacity of mesenchymal stromal cells (MSCs) cultured as monolayers. These types of two-dimensional (2D) studies can be used as simplified models to understand cell processes related to stem cell sensing and mechano-transduction in a three-dimensional (3D) context. For several other mechanisms such as cell-cell signaling, cell proliferation and cell morphology, it is well-known that cells behave differently on a planar surface compared to cells in 3D environments. In classical tissue engineering approaches, a combination of cells, 3D scaffolds and soluble factors are considered as the key ingredients for the generation of mechanically stable 3D tissue constructs. However, when MSCs are used for tissue engineering strategies, little is known about the maintenance of their differentiation potential in 3D scaffolds after the removal of differentiation soluble factors. In this study, the differentiation potential of human MSCs (hMSCs) into the chondrogenic and osteogenic lineages on two distinct 3D scaffolds, additive manufactured electrospun scaffolds, was assessed and compared to conventional 2D culture. Human MSCs cultured in the presence of soluble factors in 3D showed to differentiate to the same extent as hMSCs cultured as 2D monolayers or as scaffold-free pellets, indicating that the two scaffolds do not play a consistent role in the differentiation process. In the case of phenotypic changes, the achieved differentiated phenotype was not maintained after the removal of soluble factors, suggesting that the plasticity of hMSCs is retained in 3D cell culture systems. This finding can have implications for future tissue engineering approaches in which the validation of hMSC differentiation on 3D scaffolds will not be sufficient to ensure the maintenance of the functionality of the cells in the absence of appropriate differentiation signals.

Covalent Binding of Bone Morphogenetic Protein‐2 and Transforming Growth Factor‐β3 to 3D Plotted Scaffolds for Osteochondral Tissue Regeneration

“…We chose specifically 10 days of culture to assess the effect of GF covalent binding on early cellular differentiation. hMSCs typically need a 7‐day period of culture to adhere and form their own extracellular matrix on 3D plotted scaffolds . As a negative control, POEGMA brush‐PCL scaffolds did not show cells attached, due to their antifouling properties.…”

“…hMSCs typically need a 7-day period of culture to adhere and form their own extracellular matrix on 3D plotted scaffolds. [28] As a negative control, POEGMA brush-PCL scaffolds did not show cells attached, due to their antifouling properties. Therefore, no ALP activity, cell number or GAG amount could be measured (data not shown).…”

Human Mesenchymal Stem Cells Growth and Osteogenic Differentiation on Piezoelectric Poly(vinylidene fluoride) Microsphere Substrates

“…During culture, the influence of cell–material interaction on the fate of hMSCs is not well understood. In particular, if cell culture into 3D supports compromises the cells towards one of the particular lineages [ 5 ].…”

3D screening device for the evaluation of cell response to different electrospun microtopographies

“…However, no significant differences in DNA content and ALP activity were observed. This may be due to the early time points used for the analysis, as it is known that MSCs differentiation on 3D scaffolds is typically delayed compared to conventional 2D culture plates [52]. Furthermore, osteogenic culture medium could be used to assess the influence of the different geometrical cues superimposed on the electrospun meshes.…”