Micropatterning of Phospholipid Hydrogel Layer on the Substrate for Controlled Cell Immobilization

Cell patterning plays an important role in the field of the tissue engineering owing to the influence of surrounding environment like cell-cell interaction or cell-matrix interaction. To construct a good tissue by the cell assembly, three-dimensional (3D) culturing of the cells should be examined. We focused on this, 3D patterning hydrogel layer for cell immobilization on the substrate was prepared, by photo reactive polymer and spontaneously cross-linking polymer system. We found that when the water-soluble poly (2-methacryloyloxyethyl phosphorylcholine (MPC)-co-n-butyl methacrylate (BMA)-co-p-vinylphenylboronic acid (VPBA)) (PMBV) and poly (vinyl alcohol) (PVA) mixed together in cell culture medium; they form hydrogel under mild conditions from these solutions. The mechanical properties of the PMBV/PVA hydrogel were controllable and adjusted by the change in the concentration and mixing ratio of two polymers. Cells can be immobilized by encapsulation with the PMBV/PVA hydrogel matrix with no adverse effects on the cell functions. Also, no significant interactions occurred between polymer matrix and cells. We utilized the photoreaction of PVA at the substrate to make basement of the micropatterned layer, then, by reacting to the PMBV containing cells to form a 3D cell laden micropatterned layer. As the results, the PMBV/PVA hydrogel layer was fabricated following the PVA pattern, and cells were immobilized in the hydrogel on substrate in desired hydrogel pattern. These results will give a new glimpse towards cells patterning, tissue patterning, and tissue engineering.