The Smart Multi-Well Plate (SMWP), an open technology platform for Organ-on-Chip (OoC) technology developed as part of the Moore4Medical (M4M) consortium, aims to showcase the advantages of standardization in design, manufacturing and assembly for OoC [1]. In previously presented work [2], we showed integration and characterization of piezoelectric micropumps for in-line perfusion of OoC devices. Here we present the integration and preliminary biological evaluation of three OoC devices in a SMWP prototype. This prototype, a downscaled version of the full SMWP, is constructed using stacked, predefined layers. The reservoirs of a 96-well plate are fluidically connected to integrated OoC devices and micropumps through a fluidic circuit board (FCB). A printed circuit board, assembled below the FCB, enables the electrical interfacing. The following devices were integrated in the prototype: OoC devices from Bi/ond and BEOnChip, a microelectrode array (MEA) device from MultiChannel Systems, and piezoelectric micropumps from Fraunhofer EMFT. In the OoC devices, cell culture was performed on integrated on-chip membranes. On the MEA, neuronal cells were cultured directly on the surface of the chip. For initial experiments, static cultures were performed to investigate the biocompatibility of all included materials inside the prototype. BEOnChip performed a static cell culture with skin cells (Ha- CaT) in their device. Normal cell viability and morphology was observed after 96 h and 21 days of culture using Calcein-AM/PI live/ dead staining. MCS performed static cell culture using hiPSCs-derived neurons directly cultured on the PLO/laminin-coated MEA chips. After 14 days of culture, eGFP staining showed normal cell morphology and network formation. Bi/ond performed an endothelial cell culture (HMEC-1), showing proper cell adhesion and viability in their devices. The biological experiments under static conditions showed normal cell morphology and viability in all integrated devices. In the next phase of the project, the full SMWP platform with integrated perfusion will be used for biological experimentation to generate an air-liquid interface with skin cells (BEOnChip), a perfusable MEA (MCS) and endothelial tube formation under unidirectional flow (Bi/ond). @en

Towards increased throughput and automated workflows for organs- on-a-chip, a novel high-definition electrophysiology multiwell plate is developed in the Moore4Medical project [1]. It consists of an advanced CMOS microelectrode array (MEA) chip with 16 sampling areas, each featuring 1024 electrodes [2], and of a polymeric fluidic cartridge with 16 corresponding microchambers, each connected to an integrated pneumatic peristaltic pump [3]. It can perform 16-well assays with single-cell-resolution electrical recording and integrated microfluidics. Building on an earlier prototype, we present recent developments showing a scalable integration route and excellent fluidic and electrical functionalities of the plate in an easy-to-use system. Direct bonding of the MEA chip and the cartridge was achieved using accurate glue deposition and a high-precision pick-&-place procedure, resulting in 16 well-aligned, sealed microchambers on top of the MEA. This method is developed for high-volume production. To run the plate, a user-friendly toolbox, which can be placed inside an incubator, is made together with a data acquisition unit, a pneumatic control unit, and custom software. The plate can be clamped directly inside the toolbox, ensuring communication of the MEA with the data acquisition unit by a pogo-pin connection. The pneumatic control unit actuates the 16 integrated pumps simultaneously via three pressure lines. Hence, 16 assays can run in parallel, with simple pipetting steps for cell seeding and media perfusion. To demonstrate the plate’s functions, all the 16 microchambers were primed and filled with water and 1% PBS buffer sequentially. No liquid leakage or bubble entrapment was observed. The voltage scan of the 16 MEA areas showed uniform signals and a clear difference between the two liquids (0.08 mVrms for water; 0.02 mVrms for PBS). Plate biological-validation is currently on-going to study the effect of drugs on electrical and contractile properties of human stem-cell derived cardiomyocytes and to demonstrate the plate potential for OoC standardization and workflow streamlining. @en

Organs-on-Chip (OoC) have been a promise of microfluidics since their early days, leading to a widespread use in research. OoC adoption by industry conversely has so far been slow, resulting in a big gap between what is developed by elite pioneers and what can be offered to a broad audience. A way to bridge this gap is the consolidation of standards for design, manufacturing and qualification of OoC, making it easier for experts in different fields to combine their expertise and creating integrated systems with higher value than the sum of the parts. To this end, here we present the Smart Multi-Well Plate (SMWP), a device based on an open technology platform and the standardized 96-well plate format. Within this format, the SMWP integrates 16 OoC devices and piezoelectric micropumps, connected by a microfluidic network and controlled by integrated electronics. Results and discussion: The SMWP allows for high modularity, with design and interfacing rules according to guidelines defined by experts from industry and academia [1]. With the SMWP, OoC devices, sensors and piezoelectric micropumps can be integrated using industrial workflows exploiting automated pick-and-placing and wire-bonding machines. The modules are fluidically connected by means of a polymeric fluidic circuit board and electrically connected by reusable and disposable printed circuits boards. These take care of both control of micropumps and sensors and transfer of power and data to the outside world. Fluidic and electric performance of the plate, including piezoelectric micropumps from Fraunhofer EMFT, were tested in combination with barrier model OoC devices from BEOnChip and Bi/ond and multi electrode array chips from MultiChannel Systems. Preliminary results of the biological validation of the three embodiments of the SMWP will also be presented. Conclusions: An autonomous standardized OoC platform based on an open technology was realized by leveraging industrially upscalable processes and integrating OoC devices. @en