Program at a Glance 18.05.2021
Probing the pH microenvironment in 3D cell culture systems with high spatio-temporal resolution
Institute of Nanotechnology, National Research Council (CNR-NANOTEC) --- ITALT
Monitoring the formation of a tissue and its metabolic variations during cell culture is still a challenge and mostly limited to bulk volumetric assays. Here, we present 3D scaffolds embedding capsules-based optical sensors and demonstrate the potential of these integrated systems by calibrating and mapping in space and time pH variations of human mesenchymal stromal cells (hMSCs) in different areas of the constructs. The research leading to these results received partial funding from the European Research Council (ERC) under the European Union's Horizon 2020 research and innovation programme (grant agreement No 759959, ERC-StG 'INTERCELLMED').
Towards in-silico approaches for guiding in-vitro model experiments
National Institute for Research in Digital Science and Technology (INRIA) --- FRANCE
Spheroids, organoids and lab-on-chip culturing technologies are rapidly advancing and will become a new standard in in-vitro testing. These techniques will bring new challenges with regard to their design and implementation. We propose here realistic computer simulations techniques in which every cell is modeled along with it its bio-chemical and physical properties, including variability. Our multi-cellular models can simulate virtual experiments based on process parameters that reflect those in the real experiment. By running massively parallel and fast simulations we aim to help understanding multicellular dynamics and guide/accelerate experimental design to reduce costs.
Get to know the unique 3D cell culture technology to boost your stem cell research
OLS OMNI Life Science --- GERMANY
In recent times, the emergence of 3D cell models holds a great promise as an in vitro disease model, imitating the in vivo microenvironment. The CERO 3D Incubator and Bioreactor is a novel 3D cell culture technology developed to boost the research in stem cells, spheroids, organoids, and tissues. It is easy to operate and allows the generation of highest yields of homogeneous cells. The highest levels of homogeneity and viability of organoids and spheroids in long term cultures are just two of many benefits offered. 1-4 individually controlled CEROtubes provide the highest biomass yields in a standardized way.
Human Vascularized Immuno Bone Minitissue as Antimetastatic Drug Screening Platform
Regenerative Medicine Technologies Laboratory, Ente Ospedaliero Cantonale --- SWITZERLAND
Advanced 3D in vitro models could become relevant assets for research and pharma industry to discover new drugs against breast cancer metastasis, overcoming limitations of current in vitro models. We developed a highly complex vascularized metastatic immuno bone minitissue (MIBm) by embedding up to five human bone cell types in a 3D fibrin gel. We treated MIBm and a breast cancer cell monoculture minitissue with two FDA-approved drugs. We showed that the drug effects were strictly dependent on the presence of the bone-tumor microenvironment and we could investigate drug influence on other cell components suggesting that our MIBm could better perform antimetastatic drug screening than more simplistic models.
Towards endothelial profiling in PDMS-based microfluidic models
Jagiellonian Centre for Experimental Therapeutics (JCET) --- POLAND
Endothelial cells, lining all vessels in the body, accomplish several functions like the regulation of vascular tone and the maintenance of blood fluidity. Endothelial dysfunctions are hallmarks of several cardiovascular disease. The balance is maintained through complex processes, which are sensitive to different parameters, and is therefore essential to create models that allow to focus on relevant features. In this presentation a PDMS-based microfluidic device will be shown, which allow to mimic the in vivo conditions of 3D cylindrical structure of vessels and shear stress to which endothelial cells are exposed.
Dorplet Micoarray - an novel minaturized platform for generation of spheroids in hanging droplets
Aquarray GmbH --- GERMANY
The Droplet Microarray is a revolutionary miniaturized screening platform which allows formation of single spheroids in hanging droplets. The Droplet Microarray is a transparent and completely planar array consisting of hydrophilic spots created on a superhydrophobic background. The extreme contrast between the wettable and non-wettable areas on a standard microscopy glass slide leads to spontaneous formation of arrays of homogeneous nanoliter-sized droplets. Due to its flexibility in spot size the consumption of cells and compounds is kept to a minimum compared to 96- and 384-well plate.
Development of a chemically defined three-dimensional matrix for the expansion of pancreatic organoids using the 3-D Life Hydrogel System
Cellendes GmbH ---GERMANY
The 3-D Life Hydrogel System was used to develop a GMP-compliant matrix based on a chemically defined composition in which pancreatic organoids can be expanded. The yield and quality of organoids was optimised by testing several biomimetic compositions of hydrogels that included various extracellular matrix protein-derived peptides and increasing concentrations thereof, hydrogel stiffness, the use of matrix metallo protease cleavage sites and the modification of hyaluronic acid to become a functional and integrated part of the hydrogel system. The maintenance of the progenitor status as a central requirement of the expansion was shown by marker analyses over several culture passages.
Novel BioSettlers for gentle concentration, washing and harvesting of 3D organoids
Sudhin Biopharma Co. --- UNITED STATES
BioSettlers are closed sterile modular continuous-flow single-use disposable devices that separate cells and particles based on their sedimentation velocities. We have previously demonstrated that these devices can retain and recycle the larger (>12 microns) live and productive mammalian (CHO) single cells to a bioreactor while removing the smaller dead cells and cell debris at a controlled low flow rate. We have now demonstrated that we can retain and recycle larger (>100 microns) beads, as a proxy for 3D organoids, while removing the smaller. These BioSettlers are useful for concentrating, washing, and harvesting 3D cell culture organoids.
Monodelphis domestica - derived neurospheres: new mammalian 3D in vitro cell model for CNS development and regeneration studies
Department of Biotechnology, University of Rijeka --- CROATIA
The grey short-tailed opossum (Monodelphis domestica) provides an excellent opportunity to investigate mammalian central nervous system (CNS) development and regeneration. Indeed, M. domestica neonates can fully functionally recover their spinal cord after injury during the first two postnatal weeks. We have recently established primary dissociated neuronal cultures from both spinal cord and cortex at two postnatal days: P3-5 and P16-18, respectively. These primary cultures are able to form neurospheres that posses both neurogenic and gliogenic in vitro potential. Moreover, long-term cultures after more than 3 weeks in vitro spontaneously form organoid-like aggregates. M. domestica therefore offer a novel and robust in vitro platform for a variety of comparative and developmental studies as well as new insights into CNS regeneration..
Functionalization of hydrogels with PODS® growth factors for sustained release in 3D cell culture
Cell Guidance Systems --- UNITED KINGDOM
POlyhedrin Delivery System (PODS®) is a sustained release protein delivery system based on incorporating cargo proteins into a polyhedrin crystal lattice. In cell cultures, provision of growth factors as PODS® rather than soluble proteins can reduce costs and time. Combining PODS® and hydrogels for 3D cell culture has the potential to enhance cellular behaviour and responses whilst reducing input costs in an area where upscale of production and specificity of cell function is important. PODS® have been effectively combined with multiple hydrogel materials, demonstrating growth factor release and bioactivity. Applications include microcarriers, bioprinting, and growth factor localisation to establish gradients.
Preclinical human glioblastoma models: Advances and challenges for clinical translation
Institute of NeuroPhysiopathology, Aix-Marseille University --- FRANCE
Fifteen years after the establishment of the Stupp protocol as the standard of care to treat glioblastomas, no major clinical advances have been achieved and increasing patient's overall survival remains a challenge. Nevertheless, crucial molecular and cellular findings revealed the intra-tumoral and inter-tumoral complexities of these incurable brain tumors, and the essential role played by cells of the microenvironment in the lack of treatment ecacy. Taking this knowledge into account, fulfilling gaps between preclinical models and clinical samples is necessary to improve the successful rate of clinical trials. Since the beginning of the characterization of brain tumors initiated by Bailey and Cushing in the 1920s, several glioblastoma models have been developed and improved.We focused on the most widely used 3D human glioblastoma models, including spheroids, tumorospheres, organotypic slices, explants, tumoroids and glioblastoma-derived from cerebral organoids. We discuss their history, development and especially their usefulness.
A new three-dimensional (3D)-intestinal model
1.Institut Pasteur, Unite d'Analyse d'Images Biologiques, Paris, France. 2.Instituto Mexicano del Seguro Social, Unidad de Investigacion Medica en Medicina Reproductiva, Ciudad de Mexico, Mexico. --- MEXICO
A newly build three-dimensional (3D)-intestinal model was able to replicate the general characteristics of the human colon. This 3D-scafold system consists of, a lamina propria-like compartment made up of collagen type I as well as, THP-1 and CCD-18-Co cells. Over this basement were loaded enterocyte-like cells (Caco-2/TC7) and mucus-producing cells (HT29-MTX) which develop tight and adherens junctions and secret a mucus layer. By means of microscopy imaging, omics assays and evaluation of immune responses, was determined a very dynamic interaction between Entamoeba histolytica and the 3D-intestinal model recalling the first stages of amebiasis.
Medium recirculation using a pressure-driven flow controller for microfluidic 3-D cell culture
Elvesys --- FRANCE
This demonstration will show how to set up and control recirculation of medium over cells in microfluidic culture. An Elveflow pressure-driven flow controller and pair of 3/2 valves will be connected live to a microfluidic chip. Fluid flow control using this configuration features automated switching between two reservoirs, pumping from the fuller reservoir. Liquid exchange between reservoirs will be demonstrated in a live setup. Advantages of this platform using pulseless, stable flow control and medium recirculation for applications in 3-D microfluidic cell culture will be highlighted.
Tuneable synthetic matrices to provide physiologically relevant 3D cultures for in vitro studies
Manchester BIOGEL --- UNITED KINGDOM
3D cell culture is an increasingly reliable method to mimic the in vivo environment in vitro; however, some widely used biomaterial scaffolds have limitations as they are animal derived and lack tuneability and reproducibility. Recent advancements in synthetic tuneable peptide hydrogels have shown potential to overcome these limitations by better simulating tissue microenvironments, allowing the generation of more physiologically and clinically relevant data. Here we will demonstrate the use of such systems for the growth of 3D organoids, tumour models, and their applications more broadly within regenerative medicine and drug discovery.
Humanized 3D melanoma skin model for physiological studies of tumor aggressiveness
Instituto Tecnologia Química e Biológica --- PORTUGAL
Cutaneous melanoma is the most aggressive skin cancer with increasing incidence worldwide. Actually, for metastatic melanoma patients the prognosis remains poor. In this context, it is urgent to develop efficient therapies to treat or to prevent melanoma metastases. Lately, a strong investment has been devoted for the development of in vitro human models that mimic a better tumor environment. Here, an innovative melanoma skin model is presented, to study the underlying mechanisms affecting melanoma progression and invasion. This model allows a better understanding of tumor aggressiveness, which could reveal new potential therapeutic targets and reduce the need for animal testing.
Explore the VITVO® Experience for 3D Cell Culture
Rigenerand srl --- ITALY
Cancer research, drug discovery, cell and gene therapies development and precision medicine applications require in vitro models able to generate highly predictive and valuable data. VITVO® is a closed, portable and handheld bioreactor for 3D cell culture that can be loaded with tumor and/or normal cells in combination to rapidly recreate in vitro an in vivo-like environment.The use of VITVO® in several assays focusing on oncology and evaluating the efficacy of chemotherapy, biologics, and cell-based anti-cancer agents will be discussed in order to obtain fast read-outs and reliable data for pre-clinical investigations in drug testing, gene therapy and immuno-oncology.
A versatile microelectronic lab-on-a-chip device to model biological barriers
Biological Research Centre, Szeged --- HUNGARY
Lab-on-a-chip devices are important tools to study biological barriers leading to a deeper understanding of their physiological functions, transport mechanisms, drug delivery and pathologies. Our PDMS-based sandwich-structure microelectronic biochip enables the complex visualization of the co-culture of multiple cell types of epithelial and endothelial cell-based barriers. It incorporates automatic feeding, constant fluid flow of culture medium, the assessment of barrier integrity with permeability studies and transcellular electrical resistance evaluation with gold electrodes. Surface glycocalyx investigations using streaming potential & zeta potential measurements including morphological studies and complex RNA sequencing leads to the understanding of key surface molecule patterns and functions.
Bioengineering a 3D ECM-like hydrogel for breast organoids
INEB/i3S - Bioengineered 3D Microenvironments --- PORTUGAL
Organoids have emerged as physiologically relevant in vitro models to study breast cancer. Still, they are commonly assembled in extracellular matrix (ECM)-derived 3D matrices, such as typeI-collagen or MatrigelTM which possess poorly tunable biochemical/mechanical properties, high batch-to-batch variability and intrinsic bioactivity. In vitro models based on ECM-mimetic hydrogels, like ultra-pure alginates, present key advantages: low batch-to-batch variability; well defined and xeno-free composition; and precisely customizable biochemical/physical properties. We have shown that bioengineered alginate hydrogels support mammary epithelial cell morphogenesis into organoids. We were also able to switch in situ the viscoelastic properties of these cell-laden hydrogels, recapitulating the dynamic mechanical changes of the ECM during tumor progression.
TissueSpec® Extracellular Matrix Substrates for 3D Cell Culture
Xylyx Bio, Inc. --- UNITED STATES
The extracellular matrix (ECM) is a critical regulator of cell function with physical and biochemical properties that are specific to each tissue. Conventional in vitro models lack the tissue-specific ECM molecules and mechanics of the cellular microenvironment and thus fail to be predictive. TissueSpec® ECM substrates provide the physiologic 3D microenvironment for cells, resulting in more predictive in vitro models enabling the modulation of gene expression, proliferation, differentiation, and migration.
IVTech: advanced cell culture models to study the human metabolism
IVTech srl --- ITALY
IVTech develops advanced cell culture chambers to overcome the limitation of 3D static models. The use of these products in the study of metabolism has provided several applications in permeability studies, simulation of human digestion as well as obesity studies. The end users have developed several 3D, dynamic and interconnected in-vitro models (5D in-vitro models), overcoming the limitations related with the use of standard technology. This presentation will introduce the technology and will provide some details about how to implement an advanced in-vitro model to study the human metabolism.
SecretCells, a versatile and ergonomic microfluidic tool for secreted biomarkers monitoring from perfused 3D cell cultures
Nanobiose --- FRANCE
SecretCells® is a versatile and ergonomic microfluidic tool for 2D/3D perfused cell cultures, control cells exposure to tested items and monitoring of secreted biomarkers, in the same experiment. Created for in vitro efficacy/safety assays, SecretCells® is the combination of a microfluidic chip for cell culture, a removable biosensor to capture on-line up to 10 secreted biomarkers on a microarray and a reagent kit, to evaluate those biomarkers. The 'open' architecture of the biochip allows to assay cells both in 2D or 3D, with different types of membranes/scaffolds/matrices but also explants and biopsies, all being retrievable at endpoint for further analyses.
Magnetic 3D Cell Culture Methods and Assays
Greiner Bio-One --- UNITED STATES
Magnetic 3D cell culture (M3D) is a platform for 3D cell culture that escapes many of the limitations of current methods. The basis of this platform is the magnetization of cells and subsequent 3D culture assembly with magnetic forces. This demo will describe M3D in detail and its assays, including wound healing and high-throughput screening assays. Overall, M3D is a 3D cell culture platform that is rapid, simple to use, and scalable, making it ideal for a wide range of applications.