Fig 1. The methodology for PEGDA-based micropatterning on methacrylated glass-bottomed petri dish by photo crosslinking.

(a) Schematic illustration of the fabrication process (i) Glass surface methacrylation; (ii) PEGDA pre-hydrogel solution was added onto the glass surface, and then exposed to UV light (wavelength = 405 μm) with the virtual photomask embedded; (iii) Micropatterned PEGDA gel was fabricated on the glass surface. Uncrosslinked pre-hydrogel solution was removed by multiple PBS washes. (b) Schematic illustration of hESC seeding and culture (i) Laminin coating was applied on the micropatterned surface. Unattached laminin was washed away after 2.5-hr-incubation at 37°C; (ii) hESC suspension was added at desired density on the laminin coated surface. Unattached cells were washed away after 45-min-incubation at 37°C. Following seeding, media with desired reagents was added to the dish for further culture or induction.