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Fibroblastic reticular cells (FRCs), the T-cell zone stromal cell subtype in the lymph nodes, create a scaffold for adhesion and migration of immune cells, thus allowing them to communicate. Although known to be important for the initiation of immune responses, studies about FRCs and their interactions have been impeded because FRCs are limited in availability and lose their function upon culture expansion. To circumvent these limitations, stromal cell precursors can be mechanotranduced to form mature FRCs. Here, we used a library of designed surface topographies to trigger FRC differentiation from tonsil-derived stromal cells (TSCs). Undifferentiated TSCs were seeded on a TopoChip containing 2176 different topographies in culture medium without differentiation factors, then monitored cell morphology and the levels of ICAM-1, a marker of FRC differentiation. We identified 112 and 72 surfaces that upregulated and downregulated, respectively, ICAM-1 expression. By monitoring cell morphology, and expression of the FRC differentiation marker ICAM-1 via image analysis and machine learning, we discovered correlations between ICAM-1 expression, cell shape and design of surface topographies and confirmed our findings by using flow cytometry. Our findings confirmed that TSCs are mechano-responsive cells and identified particular topographies that can be used to improve FRC differentiation protocols.

Polystyrene (PS) is the most commonly used material in cell culture devices, such as Petri dishes, culture flasks and well plates. Micropatterning of cell culture substrates can significantly affect cell-material interactions leading to an increasing interest in the fabrication of topographically micro-structured PS surfaces. However, the high stiffness combined with brittleness of PS (elastic modulus 3-3.5 GPa) makes high-quality patterning into PS difficult when standard hard molds, e.g. silicon and nickel, are used as templates. A new and robust scheme for easy processing of large-area high-density micro-patterning into PS film is established using nanoimprinting lithography and standard hot embossing techniques. Including an extra step through an intermediate PDMS mold alone does not result in faithful replication of the large area, high-density micropattern into PS. Here, we developed an approach using an additional intermediate mold out of OrmoStamp, which allows for high-quality and large-area micro-patterning into PS. OrmoStamp was originally developed for UV nanoimprint applications; this work demonstrates for the first time that OrmoStamp is a highly adequate material for micro-patterning of PS through hot embossing. Our proposed processing method achieves high-quality replication of micropatterns in PS, incorporating features with high aspect ratio (4:1, height:width), high density, and over a large pattern area. The proposed scheme can easily be adapted for other large-area and high-density micropatterns of PS, as well as other stiff and brittle polymers. (C) 2016 Elsevier B.V. All rights reserved.

Stem cells respond to the physicochemical parameters of the substrate on which they grow. Quantitative material activity relationships - the relationships between substrate parameters and the phenotypes they induce - have so far poorly predicted the success of bioactive implant surfaces. In this report, we screened a library of randomly selected designed surface topographies for those inducing osteogenic differentiation of bone marrow-derived mesenchymal stem cells. Cell shape features, surface design parameters, and osteogenic marker expression were strongly correlated in vitro. Furthermore, the surfaces with the highest osteogenic potential in vitro also demonstrated their osteogenic effect in vivo: these indeed strongly enhanced bone bonding in a rabbit femur model. Our work shows that by giving stem cells specific physicochemical parameters through designed surface topographies, differentiation of these cells can be dictated. (C) 2017 Elsevier Ltd. All rights reserved.