A high-throughput microfluidic microphysiological system (PREDICT-96) to recapitulate hepatocyte function in dynamic, re-circulating flow conditions

Abstract

Microphysiological systems (MPSs) are dynamic cell culture platforms that incorporate mechanical and chemical cues to recapitulate organ‐specific functions and architecture. Recent advances in microfabrication technologies and processes have enabled the development of enhanced in vitro models that capture human physiology more accurately while increasing throughput and reducing material‐based technical artifacts. A common candidate for MPS development is liver due to its crucial role in drug metabolism, inflammation and human disease. While several liver‐on‐chip models exist, few studies have combined long term culture with dynamic inputs, such as media flow, in a high‐throughput format. Herein, we have developed an oxygen permeable, high‐throughput (96‐well form factor), thermoplastic MPS (PREDICT‐96 array) integrated with an on‐board, ultra‐low volume, recirculating pumping system (PREDICT‐96 pump) to facilitate media flow in the system; collectively, these technologies comprise the PREDICT‐96 platform. To demonstrate usability of the PREDICT‐96 array, we introduce PHHs using standard handling procedures for multi‐well plates and maintain viable culture for up to 14 days. By incorporating media flow to mimic in vivo mass transport, we observe a robust increase in metabolic and secretory functions of PHHs when compared to static culture. The PHHs demonstrate reproducible baseline metabolic activity and secretion markers which underscores the utility of the system for drug screening applications. Furthermore, long‐term culture with fluid flow allows for the periodic introduction of media components (e.g., fatty acids, cytokines) and resolve cellular responses to chronic stimuli over time. The low‐volume footprint of the pump and small media volume in the MPS allow for the interrogation of hepatic responses incorporating secretion feedback to a stimulus, which is essential for disease model development and drug interrogation. We envision future development of this liver model to incorporate key primary hepatic cells, multi‐cellular co‐cultures and integration with high‐throughput analytical tools. This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

High-Throughput Human Primary Cell-Based Airway Model for Evaluating Influenza, Coronavirus, or other Respiratory Virusesin vitro

“…The PREDICT96 organ-on-chip platform consists of a microfluidic culture plate with 96 individual devices and a perfusion system driven by 192 microfluidic pumps integrated into the plate lid [25,26]. Here the platform is adapted to support culture of barrier tissues, such as airway tissues, by establishment and maintenance of an ALI.…”

“…The microfluidic stack is attached to a modified 384well COP plate (Aurora Microplates, Whitefish, MT, USA) using a 0.135 mm thick pressure sensitive adhesive consisting of a 25.4 µm polyester carrier film with MA-69 acrylic medical grade adhesive ARcare® 90106 (Adhesives Research, Inc., Glen Rock, PA, USA), which had been previously laser-cut in the pattern of the well plate grid and laminated with a hydraulic press at 1.0 MPa for 2 min. Further details on the fabrication process are found elsewhere [25,26].…”

“…As described in our previous work [25], in order to establish recirculating flow in the microchannels, we incorporate a self-priming micropump array into the lid that serves as the fluidic interface with the culture plate via stainless steel tubes. The PREDICT96 pumping system has 192 individual pneumatically-actuated micropumps embedded in the plate lid: two per culture device, one for the chamber above the membrane and one for below.…”

“…These limitations include low-throughput and complex operation, a lack of relevant metrics for system assessment, the use of research-grade materials and components, the lack of critical components of tissue or organ structure required to replicate infection, and most critically, an absence of confidence in the in vitro-in vivo correlation (IVIVC) for these technologies. Our group has developed systems that offer higher throughput, compatibility with existing pharmaceutical laboratory infrastructure, convenient readouts for quality control and physiological monitoring, and that are demonstrating the potential as platforms for preclinical studies across many disease areas and application domains [25,26].…”

“…In this manuscript, we build on our previous work and the work of others to develop an in vitro model with an ALI capability sufficient to support physiologically relevant epithelial differentiation such as that in gut or kidney models [25,26]. For functional proximal airway or alveolar models, human primary cells should be cultured at an ALI representing a suitable analogue for the respiratory barrier tissues that mimics mucus formation, mucociliary flow, and a range of physiologically relevant responses [23,24].…”