Main Article Content
3D culture of stem cells has gained importance in the recent years as it is more accurate in recapturing the physiological microenvironment. Among the different methods for simulating a 3D culture microenvironment, organoid culture has a variety of applications ranging from drug discovery to miniaturized in vitro organ models. In this scenario, controlled generation of spheroids from single cells ensures consistent results especially when the spheroids are used in biosystems that require high throughput such as drug screening assays. The current study develops an efficient strategy for generation of organoids in hydrogel microwells of customizable shape and size fabricated using 3D printed micropillar templates. The efficacy of the hydrogel microwells in aggregating human pluripotent stem cells (hPSCs) is demonstrated in two different cell lines with consistent shape and size parameters. In addition to pluripotent cells, the microwells have also been shown to be effective in aggregating hPSC derived pancreatic endocrine-like cells. The versatility of this method is further demonstrated by designing tissue constructs of various non-spherical geometry, employing a hierarchical aggregation strategy in the designed microwells. Such non-spherical 3D tissue constructs is expected to more closely reproduce the biophysical attributes of the native organ.