Organoids and Spheroids Europe 2023 Agenda

Patient-Derived Organoids at the Forefront of Human Disease Modeling and Therapeutics Development
Robert Vries, CEO, Hubrecht Organoid Technology (HUB), Netherlands

TranSphere: Transporting Live Cell Cultures Under Ambient Conditions Using Organ-on-a-Chip Technology
Peter Ertl, Professor of Lab-on-a-Chip Systems, Vienna University of Technology, Austria

Living cell cultures are widely using in basic to applied research and industrial production. To meet the growing demand for living human cells in biological and medical research as well as pharmaceutical development, cell lines are either ordered and shipped from well-known cell banks around the world or sent between different laboratories. The cryogenic and live transport of cells is not only problematic since high cell viability must be guaranteed, they are also important cost factor considering the prize dry ice and portable incubators. To overcome these limitations, we have developed a microfluidic multi-spheroid array that utilizes the resilience of 3D cell aggregates to cope with physical, chemical and environmental influences such pH changes, mechanical stress and ambient temperatures. In this presentation biochip design, fabrication and initial testing results will be presented.

Human Organoid Technology to Study Human Viral Infections
Dasja Pajkrt, Professor of Viral Pediatric Infectious Diseases, Amsterdam University Medical Center, Head OrganoVIR Labs, Netherlands

Virus research historically relies on research using cell lines or animal models. Organoid technology is highly applicable in the virology field, yet unexplored. Organoid systems can mimic the in vivo human physiological environment and provide tools to study human host-virus interactions. The pathogenesis of a variety of human viruses, such as picornaviruses, HIV and human cytomegalovirus (CMV) is increasingly being studied using these novel human organoid models. Human airway epithelium (HAE) cultures and lung organoids allow for host-pathogen interaction studies on viral infections in the respiratory tract (RT), while human gut and brain organoids facilitate human gastro-intestinal tract (GIT) and brain studies. We established human RT (using HAE and lung organoids) and GIT (using human gut organoids) 3D models to study virus infections such as SARS-CoV2, picornavirus, CMV infections. During the presentation I will share results that are derived from these studies.

Scaffolded Spheroids Enabled by High-Resolution 3D Printing
Aleksandr Ovsianikov, Professor, Head of Research Group 3D Printing and Biofabrication, Technische Universität Wien (TU Wien), Austria

Mesenchymal Stromal-Stem Cells Promote Intestinal Epithelium Regeneration after Chemotherapy-Induced Damage
Magdalena Lorenowicz, Head of the Advanced In Vitro Model Systems Department, Biomedical Primate Research Center, Netherlands

Allogeneic hematopoietic stem cell transplantation (HSCT) is a curative treatment for leukemia and a range of non-malignant congenital disorders. An essential component of HSCT is the conditioning regimen, consisting of chemotherapy and/or total body irradiation, administrated prior to hematopoietic cell infusion. The success of the therapy is hampered by the development of acute graft-versus-host-disease (aGvHD), in which immune cells from the donor attack healthy recipient tissue, including the liver, skin, and gut. Especially intestinal aGvHD is a life threatening complication. Data from our institution and others demonstrate rescue of ~50% of patients suffering from aGvHD with mesenchymal stromal cells (MSCs) in Phase II clinical trials. However, the underlying mechanism of MSCs contributing to steroid-resistant aGvHD treatment is still unknown. Current models in animal studies and homogeneous cell lines are highly relevant to the field, but these approaches are limited because of the high complexity of human tissues and organs. Immortalized and human primary cell lines have enabled detailed investigation of specific cell types, but do not recapitulate the cellular heterogeneity characteristics for physiological tissues. For this purpose, a suitable in vitro model is needed, which partially recapitulates the in vivo situation of GvHD patients and allows to study the mode of action of MSCs during organ/tissue regeneration in these patients. Here we developed a novel co-culture model of chemotherapy-induced small intestine damaged organoids, self-organizing 3D mini-guts derived from human stem cells, and MSCs and studied the regenerative aspects of MSC treatment on damaged intestinal epithelium by transcriptomic and proteomic analysis. Our study shows that busulfan, the chemotherapeutic commonly used as conditioning regimen before HSCT, damaged the small intestine epithelium by affecting pathways regulating epithelial mesenchymal transition, proliferation, and apoptosis. The MSCs reversed these effects by regulating genes and proteins involved in these pathways, which we also confirmed by functional evaluation of proliferation and apoptosis. Collectively, we demonstrate that our novel in vitro co-culture model is a new valuable tool to mimic the in vivo interplay between damaged intestinal epithelium and MSCs, providing a micro-physiological environment relevant to that of GvHD, and allows to study the molecular mechanism behind the therapeutic effects of MSCs in these patients.

Deployable Extrusion Bioprinting of Compartmental Tumoroids with Cancer Associated Fibroblasts for Immune Cell Interactions
Yan Yan Shery Huang, Assistant Professor, Department of Engineering, University of Cambridge, United Kingdom

Realizing the translational impacts of three-dimensional (3D) bioprinting for cancer research necessitates innovation in bioprinting workflows which integrate affordability, user-friendliness, and biological relevance. Herein, we demonstrate 'BioArm', a simple, yet highly effective extrusion bioprinting platform, which can be folded into a carry-on pack, and rapidly deployed between bio-facilities. BioArm enabled the reconstruction of compartmental tumoroids with cancer-associated fibroblasts (CAFs), forming the shell of each tumoroid. The 3D printed core–shell tumoroids showed de novo synthesized extracellular matrices, and enhanced cellular proliferation compared to the tumour alone 3D printed spheroid culture. Further, the in vivo phenotypes of CAFs normally lost after conventional 2D co-culture re-emerged in the bioprinted model. Embedding the 3D printed tumoroids in an immune cell-laden collagen matrix permitted tracking of the interaction between immune cells and tumoroids, and subsequent simulated immunotherapy treatments. Our deployable extrusion bioprinting workflow could significantly widen the accessibility of 3D bioprinting for replicating multi-compartmental architectures of tumour microenvironment, and for developing strategies in cancer drug testing in the future.