Abstract
Human mesenchymal stem (hMSCs) are defined as multi-potent colony-forming cells expressing a specific subset of plasma membrane markers when grown on flat tissue culture polystyrene. However, as soon as hMSCs are used for transplantation, they are exposed to a 3D environment, which can strongly impact cell physiology and influence proliferation, differentiation and metabolism. Strategies to control in vivo hMSC behavior, for instance in stem cell transplantation or cancer treatment, are skewed by the un-physiological flatness of the standard well plates. Even though it is common knowledge that cells behave differently in vitro compared to in vivo, only little is known about the underlying adaptation processes. Here, we used micrometer-scale defined surface topographies as a model to describe the phenotype of hMSCs during this adaptation to their new environment. We used well established techniques to compare hMSCs cultured on flat and topographically enhanced polystyreneand observed dramatically changed cell morphologies accompanied by shrinkage of cytoplasm and nucleus, a decreased overall cellular metabolism, and slower cell cycle progression resulting in a lower proliferation rate in cells exposed to surface topographies. We hypothesized that this reduction in proliferation rate effects their sensitivity to certain cancer drugs, which was confirmed by higher survival rate of hMSCs cultured on topographies exposed to paclitaxel. Thus, micro-topographies can be used as a model system to mimic the natural cell micro-environment, and be a powerful tool to optimize cell treatment in vitro.
Highlights
Human mesenchymal stem or stromal cells have a spherical morphology in vivo and display a mostly quiescent phenotype1. hMSCs are exposed to flat tissue culture polystyrene (TCP) the moment they are harvested from the human body and cultured in vitro
Depending on the type of surface topography and cell type, induced changes in cell behavior range from initiation of osteogenic differentiation of hMSCs11, adaptation of an anti-inflammatory M2 phenotype of macrophages[12,13] or tissue formation of corneal epithelial cells[14]
HMSCs mostly adhere in the valleys, and are surrounded by 10 μm high topographical features. hMSCs changed both their cell and nuclear morphology dramatically compared to the flat reference as seen after 3 days of culture (Fig. 1A)
Summary
Human mesenchymal stem or stromal cells (hMSCs) have a spherical morphology in vivo and display a mostly quiescent phenotype1. hMSCs are exposed to flat tissue culture polystyrene (TCP) the moment they are harvested from the human body and cultured in vitro. It is important to realise that most manuscripts provide detailed reports on functional phenotypes but mostly ignore these basic parameters, it is known that microfabrication platforms are able to influence these basic phenotypes, as e.g. surface structure induced shifted cell cycle distribution[18] and water flux controlled cell volume as a response to differential cell spreading[19]. In this manuscript, we set out to map surface topography induced changes in cellular state compared to hMSCs cultured on flat substrates. We followed the adaptation of hMSC phenotype within the first hours after contact up to a few days of culture, in terms of changes in cell and nucleus shape and volume, metabolism and cell cycle progression and documented a dramatic change in cell physiology over this period of time
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