Pneumatic-aided micro-molding for flexible fabrication of homogeneous and heterogeneous cell-laden microgels

Microgels are favorable for numerous applications such as drug delivery, biomaterials science and tissue engineering. Conventionally, photolithographic methods and micro-molding techniques are extensively exploited to prepare microgels; however, they are, respectively, limited to photocrosslinkable polymers and inadequate to generate serially patterned hydrogels due to the static nature of utilized molds. Herein, we proposed a simple and versatile approach, termed pneumatic-aided micro-molding (PAM), to flexibly fabricate microgels with precise control over multiple cell types and microarchitectures of hydrogels through strategically designed pneumatic microvalves. Using the PAM approach, different cells were encapsulated in various hydrogels that had well-defined geometries. Additionally, single/multiple micro-channeled cell-laden microgels were fabricated, of which the shape, number and arrangement could be finely tuned by varying microvalve configurations. Moreover, multi-compartmental microgels comprising composite hydrogel structures were engineered following a two-step PAM, which demonstrated the utility for biomimetically constructing a three-dimensional (3D) liver microtissue composed of a radially orchestrated network of hepatic cords and sinusoids. The resulting microtissue resembled the organizational complexity of the liver lobule and was applied for the evaluation of acetaminophen-induced hepatotoxicity. Collectively, the PAM strategy could be a useful and powerful tool in biomedical engineering, in vitro 3D cell culture, and fundamental biological studies.