High resolution stereolithography fabrication of perfusable scaffolds to enable long-term meso-scale hepatic culture for disease modeling

Abstract

Microphysiological systems (MPS), comprising human cell cultured in formats that capture features of the three-dimensional (3D) microenvironments of native human organs under microperfusion, are promising tools for biomedical research. Here we report the development of a mesoscale physiological system (MePS) enabling the long-term 3D perfused culture of primary human hepatocytes at scales of over 106 cells per MPS. A central feature of the MePS, which employs a commercially-available multiwell bioreactor for perfusion, is a novel scaffold comprising a dense network of nano- and micro-porous polymer channels, designed to provide appropriate convective and diffusive mass transfer of oxygen and other nutrients while maintaining physiological values of shear stress. The scaffold design is realized by a high resolution stereolithography fabrication process employing a novel resin. This new culture system sustains mesoscopic hepatic tissue-like cultures with greater hepatic functionality (assessed by albumin and urea synthesis, and CYP3A4 activity) and lower inflammation markers compared to comparable cultures on the commercial polystyrene scaffold. To illustrate applications to disease modeling, we established an insulin-resistant phenotype by exposing liver cells to hyperglycemic and hyperinsulinemic media. Future applications of the MePS include the co-culture of hepatocytes with resident immune cells and the integration with multiple organs to model complex liver-associated diseases

Human disease models in drug development

“…Nonetheless, human-based models have so far been primarily used for toxicity testing 61 – 64 , and their application in preclinical and clinical research is still in its infancy. Promising disease models of the liver 65 , 66 , pancreas 67 , 68 , central nervous system 35 , 69 , 70 , skin 71 , lung 72 , 73 , intestine 72 – 76 , musculoskeletal system 77 and heart 78 – 81 have been developed, but their equivalence or superiority over their animal models needs to be demonstrated. In this section, successful examples of preclinical and clinical implementation of human-based disease models are discussed.…”

“…Organs-on-chips should include the same cell types with the correct proportion as real organs, such that the assembled layout contains specific cell types mixed together and others segregated into layers. For example, a liver-on-a-chip should use a physiological ratio of hepatocytes, liver sinusoidal endothelial cells, stellate cells and Kupffer cells, and layer them to mimic the morphology of the liver sinusoid 66 , 182 , 183 .…”

Bioprinting Technologies and Bioinks for Vascular Model Establishment

“…The printer only needs a movable platform in the z-axis direction, improving the printing efficiency with a sufficiently high spatial resolution. SLA has been used to fabricate a variety of complex cell-laden structures including vascular networks [79,[89][90][91][92][93][94].…”

A similarity scaling approach for organ-on-chip devices

“…3,17,19,20 But a general framework for scaling remains elusive, and recent reviews have consistently listed scaling as an urgent problem to be tackled. 1,8,21,22 We argue that the solution requires a shift in focus from the ad hoc needs of specific OoCs to a systematic view that accounts for the multiple factors involved. Here we propose one such framework by adapting the techniques of dimensional analysis and similarity, both classical tools in physics and engineering.…”

“…3,17,19,20 But a general framework for scaling remains elusive, and recent reviews have consistently listed scaling as an urgent problem to be tackled. 1,8,21,22…”