Figure 1 | NPG Asia Materials

Figure 1

From: Bio-inspired micropatterned hydrogel to direct and deconstruct hierarchical processing of geometry-force signals by human mesenchymal stem cells during smooth muscle cell differentiation

Figure 1

(a) Preparation and characterization of micropatterned polyacrylamide (PAA) hydrogel. To promote PAA hydrogel binding, coverslips were pretreated with APTES. (i) Thereafter, an acrylamide/bis mixture was sandwiched between the amino-silanized coverslips and a conventional transparency to ensure that the acrylamide and bis mixture was uniformly distributed on the coverslip. (ii) After the PAA hydrogel polymerized, the transparency was carefully peeled off to release the PAA hydrogel-coated coverslip. (iii) The PAA hydrogel surface was then covalently linked with sulfo-SANPAH to facilitate cross-linking of FN onto the surface. Thereafter, the PAA hydrogel was baked for a short time ~20 min at 60 °C to ensure complete removal of water. (iv) FN protein with defined geometries was then transfer-printed using microcontact printing. (v) The PAA hydrogel was rehydrated with cell culture medium, followed by cell seeding. (vi) Geometrically defined cells could be validated using phase contrast microscopy. Representative immunolabeled FN islands consisting of (b) circular, AR (c) 1:1, (d) 5:1, (e) 10:1 and (f) 15:1 micropatterns on engineered PAA hydrogel. Scale bar, 200 μm. (g) Reconstructed Z stack image of micropatterned PAA hydrogel revealed the typical thickness of PAA hydrogel used for this study.

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