Non-swellable F127-DA hydrogel with concave microwells for formation of uniform-sized vascular spheroids.

Abstract

Hydrogels with concave microwells are one of the simplest means to obtain uniform-sized cellular spheroids. However, the inherent swelling of hydrogels leads to reduced mechanical strength and thus deforms the structure of the microwells. In this study, we developed a hydrogel with microwells for formation of vascular spheroids via non-swellable di-acrylated Pluronic F127 (F127-DA), which showed higher mechanical strength than a conventional di-acrylated polyethylene glycol (PEG-DA) hydrogel. The uniform-sized vascular spheroids were spontaneously generated by human umbilical vein endothelial cells (HUVECs) and fibroblasts in the microwells. The endothelial functions of vascular spheroids were about 1-fold higher than those in two-dimensional (2D) culture, as indicated by secretion of nitric oxide (NO), prostacyclin (PGI2) and tissue factor pathway inhibitor (TFPI). Interestingly, the vascular spheroids with large diameter showed higher sensitivity to ethanol toxicity than those with small diameter, possibly due to the higher endothelial functions of large spheroids. Hence, F127-DA hydrogel with concave microwells provides a convenient way of forming uniform-sized spheroids that are useful for high throughput screening of drug/food toxicity.