EUROTOX 2016

Meet us in Seville at the 52nd European Congress of the European Society of Toxicology.


S. Eggert, F. A. Alexander Jr., J. Wiest: Real-time Spheroid Monitoring via an Automated Microphysiometer

Currently, in vitro toxicity studies rely heavily on chemical labelling and end-point assays to characterize the response of the body to newly developed compounds. While satisfactory for simple toxicity screening using planar cultures, chemical labels may affect the basal metabolic function of certain cell types. More physiologically relevant 3D cultures, such as Organs-on-Chips, are emerging as a advantageous tool for more accurate toxicology studies. However, conversion of these models into a high-throughput format is complicated by the requirement of significantly larger numbers of cells per model. Additionally, long-term studies require multiple samples for each time-point investigated, further increasing the necessary cell stock. For this reason, real-time monitoring techniques, like cellular microphysiometry, can provide a beneficial platform for studying 3D tissue cultures by screening multiple time-points using a single sample. Herein, we have developed a label-free technique for the culture and monitoring of 3D spheroids on a BioChip, which is able to measure extracellular acidification rate and oxygen consumption rate in real-time. 3D-printed microwell arrays were incorporated with porous layers into Intelligent Mobile Lab for In Vitro Diagnostics (IMOLA-IVD) BioChips to maintain spheroids. HepG2 liver cells were seeded to form liver spheroids at 1000 cells per spheroid over the course of four days. Mature liver spheroids measuring 622 microns in diameter were transferred onto newly designed chips and perfused for 36 hours prior to lysis with sodium dodecyl sulfate. The presented work serves as a candidate study for extended long-term monitoring of spheroids on the IMOLA-IVD.

F.A. Alexander J., J. Wiest: Skin-On-A-Biochip: Leveraging Cellular Microphysiometry for IMOLA-based Organ-on-a-Chip Studies

Currently, protocols that predict organ toxicity using animal testing are inaccurate at predicting toxicity in vivo and present an ethical dilemma in regards to animal welfare. The advent of more accurate 3D in vitro microphysiological systems, so-called organs-on-a-chip (OOCs), has improved our ability to probe the potential effects of drug candidates in vivo. Cellular microphysiometry systems like the IMOLA-IVD (cellasys GmbH), a microsensor array-based assaying technique, offer a solution to these issues with the ability to noninvasively monitor biological changes in real-time. A candidate 3D in vitro culture, the reconstructed human epidermis (RHE) artificial skin, was integrated with a prototype IMOLA-IVD biochip designed for online monitoring of commercially manufactured artificial skin models. Automated monitoring of an RHE in real-time can reveal time-resolved data on the toxic effect new compounds have on the epidermis. For this reason, we developed a protocol for an automated skin corrosion/irritation assay that monitors transepithelial electrical resistance (TEER) and extracellular acidification (ECAR). Metabolic signals were recorded in real-time in an incubator via a modified IMOLA-IVD system. EpiDerm RHE’s from MatTek In Vitro Life Sciences Laboratories were perfused automatically with medium via a peristaltic pump, IMOLA fluidic modules and the DALiA control software. Future work will involve developing this method into more assays that monitor other in vivo-like 3D cultures (i.e. liver spheroids) to develop a plug and play suite of IMOLA organ models for in vitro toxicity testing.

J. Wiest: Automated INVITTOX protocol # 130

The IMOLA-IVD technology monitors the extracellular acidification, cellular respiration and changes in impedance of living cells. In combination with a standard peristaltic pump, proprietary fluidic modules and control software it was possible to set up different cell based assays or organ-on-chip models. In this work we set up a configuration to transfer the cytosensor microphysiometer test method for identification of eye irritation which was validated by the European commission for Validation of Alternative Methods (ECVAM). It measures the extracellular acidification rate (EAR) of L929 mouse fibroblasts and the influence of e.g. the detergent sodium dodecyl sulfat (SDS) toward their EAR. The results show that the determination of the metabolic rate decrement by 50% (MRD50) value can be automated with the proposed set-up. Furthermore it was possible to further develop the INVITTOX protocol toward a fetal bovine serum (FBS) free assay. This simplifies the assay (no difference between seeding and low-buffered treatment medium) and excludes ethical issues related to FBS. Furthermore an increase in reproducibility is expected since inter-laboratory differences due to the problem of different personnel and different lots of FBS are overcome. The necessary adaptations of the INVITTOX protocol # 130 are described and measurements using FBS-free L929 fibroblasts are presented.

J. Wiest: Automated long-term monitoring of extracellular acidification and changes in impedance of living cells

Label-free monitoring of living cells is useful to determine e.g. delayed effects of chemicals which can not be addressed by conventional endpoint-assays. In the presented work a technology to dynamically monitor the energy metabolism of living cells is described. The IMOLA-IVD technology is introduced which monitors extracellular acidification and changes in bioimpedance of living cells. To allow long-term experiments, a fluidic system is included to supply the cells with fresh cell culture media and to add or remove the chemical compound under investigation. A short historical review on microphysiometry is given. Examples from application fields as oncology, toxicology, regenerative medicine and environemental monitoring are presented and further challenges such as data processing are highlighted. The presented technology is able to perform label-free long-term experiments on living cells and to reveal delayed effects of drugs or chemicals.

BIOGRAPHY

Press release: Printed sensors for research and development on antiviral drugs


Printed sensors for research and development on antiviral drugs

The efficacy of novel antivirals is increasingly studied with modern measuring methods such as real-time monitoring of live human cells. This helps to minimize the use of animal testing and increases the significance of the test for its relevance to patients. An interdisciplinary team of researchers including cellasys GmbH, AiCuris GmbH + Co. KG, SAUERESSIG GmbH + Co. KG and Haydale Limited, led by the Fraunhofer Institute for Biomedical Engineering IBMT, have now reached a milestone in the BMBF funded project BIOGRAPHY. For the first time, living cells have been examined with the help of printed graphene sensors.

Within this project, cellasys is responsible for the specification and characterization of the sensors. Currently, the growth behavior of living cells can be monitored electronically on the graphene structures using the IMOLA-Technology. To achieve this goal the scientists have developed a new graphene ink and have optimized the printing process. Stability and biocompatibility of the microstructures were both confirmed during the initial phase of the project starting in 2014.

cellasys’ BioChips have been used primarily in medical research. With these new advances in printing technology, they can now be manufactured at considerably lower costs. Thus, they are now suitable for industrial applications such as pharmaceutical research and development of drugs to combat viral infections. Additionally, applications in the field of comprehensive water monitoring are now conceivable. The projected completion date of this collaborative research project is in 2017, ending with a parallelization of the measurement setup and a final testing.

Further information: www.graphene-biosensors.eu

This research and development project is partially funded by the Innovate UK and the German Federal Ministry of Education and Research (BMBF) within the Framework Concept “Research for Tomorrow’s Production” (funding number 02PN2240) and managed by the Project Management Agency Karlsruhe (PTKA).

The company

cellasys GmbH offers system solutions for microphysiometry. These include services such as contract research, research & development, and production & maintenance. Furthermore cellasys works as consultants for development of applications, data analysis and data interpretation. The microphysiometric systems monitor different parameters directly from living cells. These parameters include extracellular acidification (pH), cellular respiration (pO2) and morphology (impedance). The measurements are label-free, parallel, continuous and in real-time. With the BioChip technology you can e.g. determine the efficiency of a drug outside of humans (or animals) prior to the start of the therapy.

Further information: www.cellasys.com

Top: Printed graphene on a transparent foil. Bottom: Encapsulated sensor for use in cellasys’ microphysiometric systems.

Top: Printed graphene on a transparent foil. Bottom: Encapsulated sensor for use in cellasys’ microphysiometric systems.

EMBC 2016

Visit our presentation “Automated Transepithelial Electrical Resistance Measurements of the EpiDerm Reconstructed Human Epidermis Model” at the 38th Annual International Conference of the IEEE Engineering in Medicine and Biology Society.


Automated Transepithelial Electrical Resistance Measurements of the EpiDerm Reconstructed Human Epidermis Model

Understanding the effect of exogenous substances on human skin is critical for toxicology assessment. To address this, numerous artificial models of the topmost layer of human skin, so-called reconstructed human epidermis (RhE), have been created in an attempt to produce a clear analogue for testing. Unfortunately, current testing modalities still rely on endpoint assays and are not capable of monitoring time-resolved changes in barrier function without using numerous redundant samples. In this work, a novel, time-resolved approach is realized by monitoring the transepithelial electrical resistance (TEER) of MatTek EpiDerm® reconstructed human epidermis model, utilizing an automated protocol with the Intelligent Mobile Lab for in vitro diagnostics (IMOLA-IVD).

INNOVATIV THROUGH RESEARCH

cellasys receives the seal of quality “Innovativ durch Forschung” from Stifterverband für die Deutsche Wissenschaft


 

IBCA 2016

cellasys at the congress “Impedance-Based Cellular Assays 2016”


Dear all,

nearly three years have passed since the last IBCA meeting in Budapest in August 2013 hosted by Laszlo Kohidai. It was the second meeting of this kind that brought together the growing number of researchers in the field of impedance-based cellular assays (IBCA) to share ideas or protocols and to discuss future directions of this exciting and most versatile, label-free approach of monitoring cell-based assays.

This summer, from August 9th to 12th, IBCA returns to Regensburg. As the host of IBCA2016 I would like to invite you and your colleagues who have an interest in non-invasive cell monitoring to join us and to be part of this year’s meeting. Please save the date and bookmark our website (www.ibca2016.net) for updates on the program and further announcements. Online registration will open on March 15th from the meeting webside. A list of invited speakers and details about the session titles will follow soon. The meeting will cover impedance-based cellular assays from these fields:

Cell Differentiation
Cell Migration
In Vitro Toxicology including Nanotoxicology
Signal Transduction (in particular related to GPCR)
Cells on Filter Supports
Cell Manipulation by Invasive Electric Fields (e.g. in situ electroporation)
3D Models
New and Complementing Techniques

The beautiful city of Regensburg also known as ‘the most Italian city north of the Alps’ will hopefully set the stage for another round of inspiring presentations and fruitful discussions about this most versatile way of whole-cell biosensing. Please check out the slide show about the city of Regensburg provided by the webside to see that I am not promising too much.

I look forward to welcome you in Regensburg for IBCA 2016.

Kind regards from Regensburg

Joachim Wegener

Chair of IBCA2016

ANALYTICA 2016

Meet us at analytica 2016 (Hall B2 / Both 307)!


25th International Trade Fair for Laboratory Technology, Analysis, Biotechnology and analytica Conference

 

25th International Trade Fair for Laboratory Technology, Analysis, Biotechnology and analytica Conference

ICOSE 2015

1st International Conference on Science and Engineering, Pekanbaru / Indonesia, 28.-29.09.2015.


The 1st Conference on Science and Engineering for Instrumentation, Environment and Renewable Energy 2015 themed “Science and Engineering for Enhancing Quality of Life and Ensuring a Better Future” will be held in Pekanbaru, Indonesia at the University of Riau, September 28-29, 2015. It is jointly organized by the University of Riau (UR) Indonesia, cellasys GmbH and the Technische Universitaet Muenchen (TUM) Germany.

Please finde the call for papers here

LOACWC 2015

Welcome to our presentation at the “Lab-on-a-Chip, Microfluidics & Microarrays World Congress” in San Diego / USA.


In the presented work, a candidate 3D in vitro culture, the reconstructed human epidermis (RhE) artificial skin, was integrated with a modified IMOLA-IVD biochip for online monitoring of extracellular acidification. RhE, grown on standard polycarbonate membrane culture inserts, mimic the structure and function of human epidermis. Monitoring the metabolic output of an RhE in real-time can reveal time-resolved data on the toxic effect new compounds have on the epidermis. Automatic screening can also provide a new pathway that can revolutionize the process of testing new chemicals for skin irritancy. For this reason, a protocol for an automated skin corrosion/irritation assay was developed. IMOLA Biochip-D’s were modified to perfuse cell culture inserts with medium while maintaining an air-liquid interface. Metabolic signals were recorded in real-time in an incubator via the IMOLA-IVD system. RhE from CellSystems GmbH (epiCS) on culture inserts were perfused automatically with medium via a peristaltic pump, IMOLA fluidic modules and the DALiA control software. Future work will involve developing this method into a validated irritancy and corrosivity assay following from OECD testing guideline 431 (corrosion) and 439 (irritation), by exposing the epiCS to potentially toxic agents.

EUSAAT 2015

Meet us at the EUSAAT 2015 in Linz / Austria.


cellasys has four contributions to the congress:

28-day microphysiological monitoring of human hepatocellular cells

Analysis of effects of the adoption of alternative methods on the number of animals used for eye irritation in the European Union

Reconstructed Human Epidermis (RhE) Monitoring via the IMOLA-IVD

Conversion of the draft OECD guideline for the Testing of Chemicals: The Cytosensor Microphysiometer Test Method

IEEE EMBC 2015

You are welcome to visit our invited session on “Label Free Live Cell Monitoring” during the 37th Annual International Conference of the IEEE Engineering in Medicine and Biology Society in Milan.


IEEE EMBC 2015 Session “Label Free Live Cell Monitoring”:

Martin Brischwein, Technische Universität München:
Sensor Based Microphysiometry
The capability of continuously monitoring cells and tissues in real-time for hours or days supports the value of a sensor-based microphysiometric approach. While there are widespread applications now for in-vitro settings, the use for smart, implanted devices is just beginning. The spectrum of analysed functional parameters comprises cellular morphological dynamics, metabolic activity and patterns of electric activity. An outline of a study on human tumor tissue samples gives an example of the potential benefits and challenges of in-vitro microphysiometry.

Joachim Wegener, Universität Regensburg:
Impedance Analysis of Different Cell Monolayers Grown on Gold-Film Electrodes
Impedance analysis of mammalian cells grown on planar film electrodes provides a label-free, non-invasive and unbiased observation of cell-based assays addressing the biological response to drugs, toxins or stressors in general. Whereas the time course of the measured impedance at one particular frequency has been used a lot for quantitative monitoring, in-depth analysis of the frequency-dependent impedance spectra is rarely performed. This study summarizes and validates the existing model for spectral analysis by applying it to eight different cell types from different mammalian tissues. Model parameters correctly predict the functional and/or structural properties of the individual cells under study.

Frank Alexander, cellasys GmbH:
Online, Label-Free Monitoring of Organ-On-A-Chip Models: The Case for Microphysiometry
Primarily composed of cells on a porous membrane embedded in microfluidic channels, organ-on-a-Chip (OOC) models are coming into the spotlight as an innovative, new approach to in vitro modeling. However, more work is required to understand the impact OOCs have on cellular function including basal metabolism, barrier resistance and oxygen consumption. Electrochemical sensor-based cellular microphysiometry provides a noninvasive, real-time methodology for monitoring these attribute and can be applied to develop robust, automated assays for organ toxicology. To date, few OOCs have been studied with integrated electrochemical sensors. In this presentation, we define organ-on-a-chip systems, outline which have been studied with integrated sensors, and present a novel method to study cells cultured directly on a porous membrane.

Cornelia Pfister, HP Medizintechnik GmbH:
Dynamic Monitoring of Cellular Metabolic Activity in Combination with Live Cell Imaging
We present an automated analysis of the cellular dynamic metabolic activity in combination with live cell imaging, an essential factor for understanding the fundamental cellular physiological responses. Therefore, we utilized the Intelligent Microplate Reader, a new analysis platform for marker-free cell-based assays in real-time. To demonstrate the benefit of the platform, we analyzed the relationship between various dynamic cell parameters (extracellular acidification, oxygen uptake, cell morphology, cell density and cell migration) of L929, a mouse fibroblast cell line, under the influence of sodium dodecyl sulfate. The dynamic kinetics of the monitored parameters are consistent and revealing much information about the transactions occurring in the cells.

Lazuardi Umar, University of Riau:
Application of Algae-Biosensor for Environmental Monitoring
Environmental problems including water and air pollution, over fertilization, insufficient wastewater treatment and even ecological disaster are receiving greater attention in the technical and scientific area. In this paper, a method for water quality monitoring using living green algae (Chlorella Kessleri) with the help of the intelligent mobile lab (IMOLA) is presented. This measurement used two IMOLA systems for measurement and reference simultaneously to verify changes due to pollution inside the measurement system. The IMOLA includes light emitting diodes to stimulate photosynthesis of the living algae immobilized on a biochip containing a dissolved oxygen microsensor. A fluid system is used to transport algae culture medium in a stop and go mode; 600s ON, 300s OFF, while the oxygen concentration of the water probe is measured. When the pump stops, the increase in dissolved oxygen concentration due to photosynthesis is detected. In case of a pollutant being transported toward the algae, this can be detected by monitoring the photosynthetic activity. Monitoring pollution is shown by adding emulsion of 0,5mL of Indonesian crude palm oil and 10mL algae medium to the water probe in the biosensor.

Johannes Clauss, Technische Universität München:
In-Vivo Cell and Tissue Monitoring with Active Implants
Active implant systems are becoming increasingly important in modern medicine. We describe the development of an implantable system for the monitoring of dissolved oxygen. Tissue oxygen saturation plays a leading role in many pathophysiological processes in the human body such as the growth of malignant tumors or the viability of transplanted organs. The implant allows monitoring the tissue oxygenation in vivo with a wireless interface to an external device. An improved self-calibration technique is described to minimize sensor drift with electrochemical sensors in vivo for a better long term stability of the implant system. The sensor was coated with a hydrogel membrane to avoid convection artifacts during calibration procedure.