3D-Culture, Organoids and Organs-on-Chips 2021 Agenda

Morning Break

Automated 3D Cell-Based Assays Using a Novel Flowchip System
Evan Cromwell, President & CEO, Protein Fluidics

There is an increasing interest in using three-dimensional (3D) cell structures for modeling tumors, organs, and tissue to accelerate translation research. Significant progress has been made in formation of such structures to recapitulate the in vivo environment but performing complex assays with them can be challenging.  Organoids can be subject to loss or damage from pipettes during common steps such as media exchange, supernatant sampling, or immunofluorescence staining.  We present here results from automated organoid assays using a Pu·MA System for research in oncology therapeutics, single organoid secretion, and metabolite sampling.  The system uses a novel flowchip that contains organoid samples connected to multiple reservoirs via microfluidic channels.  Assay reagents are transferred in and out of the sample well without loss of or damage to the organoids.  The bottom of the sample chamber is thin optically clear plastic compatible with high resolution fluorescence imaging.  The whole system can be kept in an incubator allowing long term cellular assays to be performed.  In a first example, effects of anti-cancer drugs on cancer spheroids were assayed using high content imaging.  Confocal imaging allowed 3D resolution of spheroid structures and complex analysis of cytotoxic effects.  In addition, conditioned media was removed and analyzed for VEGF.  In a second example, triple negative breast cancer (TNBC) patient-derived organoids were used for disease modeling.  This novel assay system using microfluidics enables automation of 3D cell-based cultures that mimic in vivo conditions, performs multi-dosing protocols and multiple media exchanges, provides gentle and convenient handling of spheroids and organoids, and allows a wide range of assay detection modalities.

Lunch Break

Tumor-on-a-Chip Platforms for Analyzing Intact Tumor Biopsies
Albert Folch, Professor of Bioengineering, University of Washington, United States of America

Cancer remains a major healthcare challenge worldwide. It is now well established that cancer cells constantly interact with fibroblast cells, endothelial cells, immune cells, signaling molecules, and the extracellular matrix in the tumor microenvironment (TME). Present tools to study drug responses and the TME have not kept up with drug testing needs. Oncology drugs typically take 10 years and cost an average of 1 billion dollars to develop. The number of clinical trials of combination therapies has been climbing at an unsustainable rate, with 3,362 trials launched since 2006 to test PD-1/PD-L1-targeted monoclonal antibodies alone or in combination with other agents. We have developed a microfluidic platform (called Oncoslice) for the delivery of multiple drugs with spatiotemporal control to live tumor biopsies, which retain the TME. We have developed the use of Oncoslice for the delivery of small-molecule cancer drug panels to glioblastoma (GBM) xenograft slices as well as to slices from patient tumors (GBM and colorectal liver metastasis). In addition, we have developed a precision slicing methodology that allows for producing large numbers of cuboidal micro-tissues (“cuboids”) from a single tumor biopsy. We have been able to trap cuboids in arrays of microfluidic traps in a multi-well platform. This work will potentially allow for the high-throughput application of drugs to intact human tumor tissues.

Organoids For Disease Modeling and In Vitro Assessment of Drug Effects by High-Content Imaging and 3D Analysis
Oksana Sirenko, Sr. Imaging Applications Scientist, Molecular Devices

3D cell models representing different tissues were successfully used for studying complex biological effects, tissue architecture, and functionality. While complexity of 3D models remains a hurdle for the wider adoption in research and drug screening, significant progress has been made in developing tools to automate the culture, monitoring, and analysis of these structures. This enabled an increase in quality of information, throughput, and precision of complex biological assays. We describe automated cell culture and high-content imaging methods that allow monitoring and characterization of growth and differentiation of lung organoids, as well as toxicity and drug effects. Increase in size and complexity of developing organoids was monitored during eight weeks of development. We demonstrated concentration-dependent toxicity effects of several drugs that have been known to cause lung damage. In addition, responses to several anti-cancer drugs were monitored in 3D tumoroids derived from primary tumor tissues.  Advanced image analysis allowed 3D reconstitution and complex analysis of organoid structures, characterization of cell morphology and viability, as well as determining expression level of differentiation markers. Image analysis included traditional and Machine Learning-based tools to increase the ability to discern important biological information from the complex multi-dimensional datasets. These quantitative characterization of organoids morphology can be used for studying disease phenotypes and compound effects in complex assay models.

Mid-Afternoon Break

Magnetic 3D Cell Culture as an Enabling Tool
Glauco Souza, Director of Global Business Development and Innovation, 3D Cell Culture at Greiner Bio-One; Adjunct Assistant Professor, Greiner Bio-One and University of Texas Health Science Center at Houston

The growing push for 3D cell culture models is limited by technical challenges in handling, processing, and scalability to high-throughput applications. To meet these challenges, we use our platform, magnetic 3D cell culture (M3D), in which cells are individually magnetized and assembled with magnetic forces. In magnetizing cells, not only do we make routine cell culture and experiments feasible and scalable, but we also gain control in the formation and manipulation of spheroids and more complex structures. This presentation will focus on recent developments using this platform. Specifically, we will present a tool for manipulating cells and phenotypic profiling of cell types within spheroids. Overall, we apply M3D to improve the development of in vitro microphysiological systems and applications.

YAMAHA CELL HANDLER: Automated, Rapid Spheroid, Organoid and Single Cell Sort System by Imaging and Picking
Yuichi Hikichi, General Manager, MDB Division , Yamaha Motor

By using various parameters of images, like size, shape and fluorescent intensity, the CELL HANDLER^TM can automatically select single cells or 3D spheroids and sort them to various types of culture plates. It uses a unique disposable tip to ensure the damage-free and accurate handling and enables direct picking of organoids from gel. Conventional manual methods have been hampered by low throughput and suboptimal accuracy. Conventional cell sorters damage cells and need significant amounts of cells and are not applicable for spheroids and organoids. The CELL HANDLER complements these deficiencies. It enables more accurate and higher throughput sorting than manual operation and low amounts of cells (e.g. 100-1000 cells) can be sorted without damages. Any types of plates (e.g. microtiter plates, micro cavity plates, 3D colony in gel, 2D colony on plate, organ-on-a-chip) can be used as a source plate and destination plate, which will support many types of applications. The CELL HANDLER is a high-speed, high-precision pick and place system coupled with image capture and analysis capability that greatly increases the efficiency and reliability of spheroid/organoid and single cell assays.

Close of Day 1 of the Conference

Morning Break

Multi-Organ Interconnected Human-on-a-Chip Systems to Develop Phenotypic Disease Models For In Vivo Predictions
James Hickman, Professor, Nanoscience Technology, Chemistry, Biomolecular Science and Electrical Engineering, University of Central Florida; Chief Scientist, Hesperos, United States of America

I will describe such systems being constructed that are guided in their design by PBPK models. They are “self-contained” in that they can operate independently and do not require external pumps as is the case with other microphysiological systems. They are “low cost”, in part, because of the simplicity and reliability of operation allowing the observation of the effects of not only drugs but their metabolites. While systems can be sampled to measure the concentrations of drugs, metabolites, or biomarkers, they also can be interrogated in situ for functional responses such as electrical activity, force generation, or integrity of barrier function. Operation up to 28 days has been achieved allowing observation of both acute and chronic responses with serum free media. We have worked with various combinations of internal organ modules (liver, fat, neuromuscular junction, skeletal muscle, cardiac, bone marrow, blood vessels and brain) and barrier tissues (e.g. skin, GI tract, blood brain barrier, lung and kidney) and immune components. The use of microelectrode arrays to monitor electrically active tissues (cardiac and neuronal) and micro cantilevers (muscle) have been demonstrated. Most importantly, these technical advances allow prediction of both a drug’s potential efficacy and toxicity (side-effects) in pre-clinical studies.

Formation of Uniform, Size-Controllable Multicellular Tumor Spheroids in Polydimethylsiloxane-Coated Well Plates
Jirina Kroupová, Researcher, University of Chemistry and Technology, Prague, Czech Republic

Lunch Break

Rapidly Deployable and Robust 3D Cell-based Assay Systems Developed to Accelerate Biomedical Research
Anthony Davies, Adjunct Assistant Professor, Trinity College Dublin, Ireland

Historically, biomedical researchers have utilized two-dimensional (2D) cell culture models to study disease and evaluate new therapeutic approaches. However, over time there has been a shift from these traditional approaches towards three-dimensional (3D) cell culture methods. This transition has brought with it an influx of technologies capable supporting the formation and growth these new 3D cellular models. Over time strong evidence is emerging that many of these 3D cell culture approaches offer a more biologically relevant output than those derived from 2D cell culture. As the use of these 3D approaches increases, so does the demand for increased throughput and scalability. Over time it has become evident that many of these technologies do not perform well in automated lab environments hence rate limiting progress. In this presentation we cover our work in the area of 3D cell culture using world’s first liquid scaffold and the very latest solid state microplate bioreactor systems have enhanced our capability to automate and scale 3D cell based assays.

3D Vascularized Lung Barrier Tissue Model in Organ-on-a-Chip Platform with an Open Access
Min Jae Song , Staff Scientist, National Center for Advancing Translational Sciences (NCATS), United States of America

NCATS tissue bioprinting lab developed a protocol for 3D vascularized lung barrier tissue models using a commercial organ on a chip platform with an open access (OrganoPlate® Graft). This open access enabled the formation of small airway or alveolar epithelium on the top of 3D vascularized construct, exhibiting comparable barrier functions. The presentation will describe this 3D lung barrier tissue on a chip including perfused vasculature with morphological and functional validations. I will further discuss the potential applications in modeling relevant diseases.

Bioprinting of 3D Extracellular Matrix Models For Cancer Research
MoonSun Jung, Senior Research Officer, Children’s Cancer Institute Australia, Australia

Cancer is a leading cause of mortality and morbidity worldwide. However, the exact molecular mechanisms underlying the tumorigenesis and metastasis are still poorly understood. Conventional 2-dimentional (2D) in vitro models are widely used for cancer research. These monolayer models however cannot accurately recapitulate the characteristics of native tumor microenvironment, including the cell-cell and cell-matrix interactions that cancer cells experience in vivo. 3D bioprinting provides an exciting opportunity to create in vitro tumor microenvironment with living cells using extracellular matrix (ECM) materials. However, it is still rare that 3D bioprinted models use biomaterials with high biocompatibility and tunability of the ECM-like properties. In collaboration with Inventia Life Science, we have developed 3D matrix models by printing small droplets of cells and matrix components using their drop-on-demand 3D bioprinter. With this technology, we are currently investigating the diverse aspects of cancer behaviours in response to environmental changes such as stiffness and biological molecules and identifying optimal biomimic ECM conditions for different cancer types.

Poster Presentation by Dr. Oksana Sirenko, Molecular Devices. Poster Title: "3D Imaging and Analysis of Angiogenesis in the Organ-on-a-Chip Platform." Download Poster PDF by Clicking Here

Poster Presentation by Dr. Oksana Sirenko, Molecular Devices. Poster Title: "Lung Organoids for Disease Modeling and Toxicity Assessment by 3D-High Content Imaging and Analysis." Download Poster PDF by Clicking Here

Poster Presentation by Dr. Evan Cromwell, Protein Fluidics. Poster Title: Multifunctional Profiling of Patient-Derived 3D Cell Models