Publication date: 25th July 2016
The geometry has emerged as an important cue that can be transduced into biochemical signals and result in cell responses. Significant efforts have been directed to understanding the geometric cues that influence stem cells behavior on 2D, however, how 3D geometry affects cell function remains elusive.
In the present study, we used photopolymerized methacrylated hyaluronic acid (MeHA) hydrogels to construct artificial single cell 3D microcompartments with a variety of shapes (cubic, cylindrical and prismatic). MeHA hydrogels with microwells were produced by photopolymerizing MeHA against a silicon master, cell-adhesion proteins could be selectively coated into microwells, the shape, size, composition and stiffness of microwells could be well-controlled as well, giving a targeted investigation of the synergy between cell shape, biophysical, and chemical signals. After encapsulated single cells in microwells, a permeable MeHA hydrogel cover was used to envelop single cells in microwells.
We found cell spreading were predominantly limited to within 3D microwells even after 48 h in culture, cells can completely filled out microwells of all different geometries with a higher degree of F-actin concentration along the edges after 48 hours culture. Myosin IIa staining indicated that cells in all microwells can generate actomyosin contractility along the edges. In cubic and prismatic shapes, we found more bright myosin IIa staining, indicating regions of local curvature on shapes can affect cytoskeletal tension of adherent cells. The focal adhesions were also predominate at the acute corners and edges in cubical and prismatic shape. In cylindrical microwells, there were less focal adhesions formation observed. We therefore conclude that our 3D hydrogel will provide a platform to culture cell and probe single stem cell fate (i.e. differentiation), and has potential applications in investigating cell-cell or cell-matrix communication in local 3D environment.
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