Figure 2.

Schematic diagram showing the preparation of the cell-laden layer-shaped alginate hydrogels. STEP 1: A filter paper soaked in the cross-linking agent (CaCl₂ or calcium gluconate) was placed on the bottom of each well of a 6-well culture plate. The alginate-hADSCs suspension was added above and covered with the second filter paper soaked in the same gelling agent. CaCl₂ or calcium gluconate was added on the top of the construct and the plate was incubated for 45–60 min at 37°C in a humidified atmosphere of 5% CO₂; STEP 2: The excess of the cross-linking agent was removed together with both of the filter papers used. The resulting thin layer hydrogel (5 mm thickness) was washed extensively with saline buffer and GCM.