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). ... Read more

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. ... Read more

Sensing flow rates in structured microenvironments like lab-on-chip (LOC) and organ-on-chip (OoC) is crucial to assess important parameters such as transport of media and molecules of interest. So far, these micro-electromechanical systems for biology (bio-MEMS) mostly rely on flow sensing systems based on thermal sensors. However, thermal flow sensing has limitations, since the measurement principle, which is based on generation of heat, can negatively affect the biological system by increasing the fluid temperature above physiological conditions. To overcome this issue, we propose a novel electro-mechanical flow sensor centered around the deformation of a cantilever made of a thin and biocompatible ionic electroactive polymer. The polymer, called ionic polymer metal composite (IPMC), is doped with ions naturally present in most cell media for LOC and OoC devices. Unlike already existing cantilever-based systems which rely on piezo sensitive materials, our IPMC-based flow sensor shows durability in wet environment. We were able to successfully measure pulsatile flow induced by pipetting with flowrate gradually increasing from 10μL/s to 40μL/s. The proposed flow sensor shows good sensing capabilities (4.78 mV/(μL/s)) with a linear behavior in the studied range. This work sets a milestone for using flexible, electroactive materials for sensing applications in delicate biological microenvironments. ... Read more

We present preliminary recordings on chip of three-dimensional (3D) electric neuronal activity from cultures of cortical neurons derived from human-induced pluripotent stem cells (hiPSCs). The recordings were obtained through 3D microelectrode arrays (MEAs) composed of truncated, 90 μm-high Si micropyramids endowed with multiple, electrically distinct, and vertically arranged TiN microelectrodes. The unique design and implementation of the 3D microelectrodes, complemented by a 60-electrode readout interface, allow for 3D spatial recording of neuronal activity, as well as single-unit recordings in high throughput, which are currently not possible with commercial MEA platforms. Future work will aim at optimizing extended 3D MEAs over optically transparent substrates for electro-physiological investigation of 3D neuronal tissues and organoids. ... Read more

Stemming from the convergence of tissue engineering and microfluidics, organ-on-chip (OoC) technology can reproduce in vivo-like dynamic microphysiological environments for tissues in vitro. The possibility afforded by OoC devices of realistic recapitulation of tissue and organ (patho)physiology may hold the key to bridge the current translational gap in drug development, and possibly foster personalized medicine. Here we underline the biotechnological convergence at the root of OoC technology, and outline research tracks under development in our group at TU Delft along two main directions: fabrication of innovative microelectromechanical OoC devices, integrating stimulation and sensing of tissue activity, and their embedding within advanced platforms for pre-clinical research. We conclude with remarks on the role of open technology platforms for the broader establishment of OoC technology in pre-clinical research and drug development. ... Read more