From Model System to Therapy: Scalable Production of Perfusable Vascularized Liver Spheroids in “Open-Top“ 384-Well Plate

Abstract

Vasculature is a key component of many biological tissues and helps to regulate a wide range of biological processes. Modeling vascular networks or the vascular interface in organ-on-a-chip systems is an essential aspect of this technology. In many organ-on-a-chip devices, however, the engineered vasculatures are usually designed to be encapsulated inside closed microfluidic channels, making it difficult to physically access or extract the tissues for downstream applications and analysis. One unexploited benefit of tissue extraction is the potential of vascularizing, perfusing, and maturing the tissue in well-controlled, organ-on-a-chip microenvironments and then subsequently extracting that product for in vivo therapeutic implantation. Moreover, for both modeling and therapeutic applications, the scalability of the tissue production process is important. Here we demonstrate the scalable production of perfusable and extractable vascularized tissues in an “open-top“ 384-well plate (referred to as IFlowPlate), showing that this system could be used to examine nanoparticle delivery to targeted tissues through the microvascular network and to model vascular angiogenesis. Furthermore, tissue spheroids, such as hepatic spheroids, can be vascularized in a scalable manner and then subsequently extracted for in vivo implantation. This simple multiple-well plate platform could not only improve the experimental throughputs of organ-on-a-chip systems but could potentially help expand the application of model systems to regenerative therapy.

Subtractive manufacturing with swelling induced stochastic folding of sacrificial materials for fabricating complex perfusable tissues in multi-well plates

“…Compared to a previous published platform, IFlowPlate, from our group, which also allows for the generation of highthroughput vascularized tissues, the AngioPlate provides users with the capability to create hierarchical vascular networks with pre-defined number of vessel branches and diameters. 34,35 AngioPlate also allows users to create perfusable epithelial tubes that don't easily self-assemble like endothelial cells. When compared to the previously published InVADE platform, epithelial tissues on AngioPlate does not contain a thick polymer wall which creates an artificial barrier that doesn't allow for mechanical actuation.…”

Implementing organ-on-chip in a next-generation risk assessment of chemicals: a review

“…RHE models are especially acknowledged for their reproducibility but they do not meet the requirement for permeation studies. Schäfer-Korting et al (2006) showed that permeation of caffeine and testosterone using RHE is overestimated compared to the human epidermis. Therefore, RHE has limitedly utility for NGRA studies that require barrier function but may represent a useful tool for corrosion, irritation and skin sensitization potentials (Zhang et al 2017a;Song et al 2018;Teimouri et al 2019;Mehling et al 2019).…”

“…Plate-based and hydrogel micromolding and bioprinting address the limitation of spheroid size variability by directing the assembly. The iFlowPlate™ by Lin et al ( 2021 ) currently offers an approach to produce scalable perfusable vascularized liver spheroids for OoC without bioprinting.…”

Beyond PDMS and Membranes: New Materials for Organ-on-a-Chip Devices

“…In the research papers section, Hosic et al report a new approach to rapidly prototype organs-on-chips via laser cutting and assembly with double-sided adhesives, a technique that could be implemented by many laboratories using relatively inexpensive equipment . Lin et al describe a scalable system for vascularization of liver spheroids in a 384-well platform . To replace PDMS in devices that mimic barrier function, Arik et al.…”

“…27 Lin et al describe a scalable system for vascularization of liver spheroids in a 384-well platform. 28 To replace PDMS in devices that mimic barrier function, Arik et al describe a viable new approach to construct membranes from collagen type I. 29 Furthermore, Zamprogno et al elaborate on the development of collagen/elastin membranes and assessment of their mechanical properties.…”